Basic Genetics Chapter 13 + 14

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What exactly is a DNA sequence of a gene?

A DNA sequence of a gene is the order or nucleotides on a strand of DNA. There are four nucleotides: adenine, guanine, cytosine, and thymine. The sequence (the order) of these four nucleotides in a chain of DNA constitutes the gene in question. Every three nucleotides code for a protein residue, so the order of these nucleotides are important, because the order of the protein residues are important. If the sequenced is altered, the gene no longer reads the same, and its functionality may be altered.

Restriction enzyme

A bacterial enzyme that cuts DNA at specific sites.

Yeast artificial chromosome (YAC)

A cloning vector that has telomeres and a centromere that can accommodate large DNA inserts and uses the eukaryote yeast as a host cell.

genetically modified organism (GMO)

A general term used to refer to transgenic plants or animals created by recombinant DNA techniques.

Probe

A labeled nucleic acid used to identify a complementary region in a clone or genome.

Polymerase chain reaction (PCR)

A method for amplifying DNA segments using cycles of denaturation, annealing to primers, and DNA polymerase-directed DNA synthesis.

Southern Blot

A method for transferring DNA fragments from a gel to a membrane filter, developed by Edwin Southern for use in hybridization experiments.

DNA microarray

A series of short nucleotide sequences placed on a solid support (such as glass) that have several uses, such as detection of mutant genes or differences in the pattern of gene expression in normal and cancerous cells.

Recombinant DNA technology

A series of techniques in which DNA fragments from an organism are linked to self-replicating vectors to create recombinant DNA molecules, which are replicated or cloned in a host cell.

DNA sequencing

A technique for determining the nucleotide sequence of a DNA molecule.

Why would biopharming myozyme not work with bacteria?

Bacteria cannot properly fold the eukaryotic protein.

DNA fingerprint

Detection of variations in minisatellites used to identify individuals.

True or False: All bacterial cells have plasmid DNA.

False

Genomic library

In recombinant DNA terminology, a collection of clones that contains all the genetic information in an individual.

How did they catch the Golden State Killer?

The DNA at the crime scene matched the killer's 3rd cousin's DNA in a geneology databsse, then genealogy family trees were traced back to a great-great-great-grandfather to find the common ancestor to the third cousin and suspect.

Biotechnology

The use of recombinant DNA technology to produce commercial goods and services.

What do restriction endonucleases do?

These enzymes break phosphodiester bonds within a specific DNA sequence.

Which product of biotechnology helps to prevent blindness in children in third world countries?

yellow rice

Induced pluripotent stem (iPS) cells

Adult cells that can be reprogrammed (induced) by gene transfer to form cells with most of the developmental potential of embryonic stem cells. Because of this developmental potential, such cells are pluripotent.

Making human proteins to treat diseases in animal and plant hosts offers several advantages over extracting those proteins from human and animal sources. What are some of those advantages?

Advantages of making human proteins to treat disease in animal and plant hosts include increased yield, increased safety, and increased ease of acquirement. For example, insulin was harvested from the organs of mammals for use as treatment for humans with diabetes. Since the advent of recombinant DNA technology, insulin has been harvested from microorganisms more easily and with higher yields and more reliability than mammal organs. A case that concerns safety and reliability improvement is the production of hemoglobin for hemophiliacs. Before microorganisms produced it, hemoglobin was harvested from donated human blood. Many times HIV was transmitted through the donated blood along with the hemoglobin, infecting hemophiliacs who accepted the donated blood. Since the advent of recombinant DNA technology, the hemoglobin harvested for hemophiliacs has not been associated with blood bourn disease.

Why are people concerned about CRISPR? Evolution is what has driven life on Earth from billions of years. CRISPR will undermine evolution with unnatural selection and nonrandom mutation. That scares me. They are Luddites and are afraid of all new technology or technology in general. All of these answers are reasons people are afraid of CRISPR. They don't know what is feels like to suffer from a genetic disease and is more afraid of someone playing "God." They are afraid of a resurgence of supremacy of the powerful and eugenics. Who decides that dwarfism is a "disease"?

All of these answers are reasons people are afraid of CRISPR.

In cloning human DNA, why is it necessary to insert the DNA into a vector such as a bacterial plasmid?

Bacteria are able to incorporate plasmids into their genome easily. When human DNA is inserted into a vector, like a plasmid, the vector can be inserted into a bacterial cell. This creates a bacterium with human DNA in its genome. The bacterium can create multiple copies of its genome, which now include the newly inserted plasmid. This allows scientist to create many cloned copies of human DNA by using plasmids and bacteria.

Restriction enzymes are derived from bacteria. Why don't bacterial chromosomes get cut with the restriction enzymes present in the cell? Why do bacteria have these enzymes?

Bacteria are not affected by their own restriction enzymes because they either do not carry the recognition sites associated with their restrictive enzymes, or the recognition sites that are located in the bacterial genome are chemically modified to not associate with the enzymes.

What are "Bt crops"? What potential risks are associ-ated with this technology?

Biotech (Bt) crops are also known as transgenic crops or genetically modified crops, such as cotton, corn and soybeans. These types of crops are resistant to insect pests. They are modified to carry a gene that produces a toxin that kills insect pests after ingestion. The gene is derived from bacteria, which codes for a toxin that kills the pest after it has entered its gut. The risk involved in these types of crops is ecological. There is a chance that the pests will evolve defense mechanism against the toxins used. This could produce super resistant pests that would out compete other species and over run an ecosystem.

With the case of the Golden State killer, which database(s) is(are) used to find a match to a crime scene DNA profile?

CODIS, military and genalology

Embryonic stem cells (ESCs)

Cells in the inner cell mass of early embryos that will form all the cells, tissues, and organs of the adult. Because of their ability to form so many different cell types, these cells are called pluripotent cells.

The creation of transgenic crop plants using recom-binant DNA methods involves the transfer of just one gene or a small number of genes to the plants, in con-trast to classical breeding methods in which hundreds or even thousands of genes are transferred at once. Explain why this is true. If fewer genes are transferred during the creation of transgenic crops, why are some people afraid that they are dangerous?

Classical plant breeding methods involve artificial selection in which crops with desired traits (such as high yield) are used to breed crop with desired traits. Classical breeding methods create crosses between plants that trade thousands of genes. Humans have been following this method for thousands of years (classical Mendelain genetics). Transgenic crop plants use recombinant DNA methods to also produce crops with desired traits. The difference is in the method of gene manipulation. Transgenic crops are created by inserting a few desired genes directly into a plant's genome at once. This is in contrast to classical plant breeding, where a cross between two plants may produce a crop with the few desired genes, but will also transfer thousands of other genes in the process. The fear of transgenic crop danger permeates throughout the media. Part of this fear may stem from a health perspective concerning a lack of understanding of the technology and method used. Another part of this fear may stem from the unprecedented preciseness of gene selection. The notion of "playing with nature" is prevalent because of its efficiency, possibly creating the notation of "its too good to be true, so it must be dangerous." Other fear stands from an ecological standpoint, arguing that gene manipulation will produce a superior crop that will eventually produce superior pests.

1. Cloning is a general term used for whole organisms and DNA sequences. Define what we mean when we say we have a clone.

Clones in this context are defined as individuals, cells, or molecules that arise from a common ancestor. Clones are usually identical, and vary due to environmental conditions. Identical twins are considered clones because they are derived from the same cell, a fertilized egg.

Briefly describe how to clone a segment of DNA. Start with cutting the DNA of interest with a restriction enzyme.

Cloning a segment of DNA is a multistep process. The first step is acquiring the segment of DNA to be cloned by the use of restriction-enzymes. A restriction enzyme will cut the DNA into pieces wherever recognition sites are found, so the proper restriction enzyme must be used to isolate the strand of DNA to be cloned. Next the same restriction enzyme is used to cut DNA plasmids into pieces. Using the same restriction enzyme for the human DNA and the plasmid DNA ensures that the sticky tails left hanging from the cut pieces of DNA will align to produce recombinant DNA pieces after they are allowed to mix. Finally, the recombinant DNA (which includes the human DNA of interest to be cloned) is inserted into bacterial cells. The bacteria cells divide, creating multiple copies of the recombinant DNA, thus cloning human DNA in the process.

DNA profiles have been used in identifying criminals, establishing bloodlines of purebred dogs, identifying endangered species, and studying extinct animals. Can you think of other uses for DNA profiles?12.

DNA finger printing has many uses which include diagnosis and treatment of inherited disease such as Huntington's disease, cystic fibrosis, and hemophilia. DNA fingerprinting can be used in disease identification by pattern analysis. Early detection of heritable diseases is possible through DNA fingerprinting. In addition to genetic disease detection, paternity and maternity lines can be established in court cases. Other uses for DNA fingerprinting are personal identification applications. Wide use of DNA fingerprinting can be used as a replacement for identification cards, debit and credit cards, and traditional fingerprints.

Put the items in the correct order. Below is a sequence of events. Place them in the order they should occur, number 1 being the first item. Select the step number from the drop down next to each item. Items to order: DNA polymerase copies a template strand of DNA by placing complementary nucleotides against the template one at a time and joining the new nucleotide to the 3' end of the adjacent nucleotide of the growing strand. DNA helicase unwinds a short segment of the DNA making two template strands. DNA primase makes a short piece of RNA to start the new strand because DNA polymerase can only add onto existing nucleic acid. The enzyme cannot start by putting down the first nucleotide.

DNA helicase unwinds a short segment of the DNA making two template strands. DNA primase makes a short piece of RNA to start the new strand because DNA polymerase can only add onto existing nucleic acid. The enzyme cannot start by putting down the first nucleotide. DNA polymerase copies a template strand of DNA by placing complementary nucleotides against the template one at a time and joining the new nucleotide to the 3' end of the adjacent nucleotide of the growing strand.

DNA sequences can reveal many important pieces of information regarding cancer-causing genes. The sequence of nucleotides in a gene that causes cancer can be compared to the sequences of nucleotides of a non-mutated gene that does not cause cancer. A researcher can pinpoint the exact type of mutation, sometimes to the nucleotide that is causing the disease. From here, he can extrapolate what might have caused the mutation, and how to avoid environmental factors that might antagonize it. If the gene that causes disease is not mutated, but inherited, a researcher can use sequence data to determine who is at risk for disease development using the known sequence to compare it with.

DNA sequences can reveal many important pieces of information regarding cancer-causing genes. The sequence of nucleotides in a gene that causes cancer can be compared to the sequences of nucleotides of a non-mutated gene that does not cause cancer. A researcher can pinpoint the exact type of mutation, sometimes to the nucleotide that is causing the disease. From here, he can extrapolate what might have caused the mutation, and how to avoid environmental factors that might antagonize it. If the gene that causes disease is not mutated, but inherited, a researcher can use sequence data to determine who is at risk for disease development using the known sequence to compare it with.

Place the following steps of PCR in order. Extension step Cycling step Denaturing step Annealing step

Denaturing step Annealing step Extension step Cycling step

True or False: DNA profiling can definitively catch the guilty - criminal or absent father.

False

Clones

Genetically identical molecules, cells, or organisms, all derived from a single ancestor

You are running a PCR to generate copies of a fragment of the cystic fibrosis (CF) gene. Beginning with two copies at the start, how much of an amplification of this fragment will be present after six cycles in the PCR machine?

In this problem, the amount of fragments present in the 6th round would be 64. Each polymerase chain reaction doubles the amount of fragments there are. So at the end of each round, double the amount of fragments are present than were the previous round. A mathematical shortcut of this calculation would use exponents. The base would represent the original amount of fragments, and the exponent would represent the round we would like to calculate. In our case, 2 for the original amount of fragments and 6 for the number of PCR rounds, so 26=64. The following is a visual representation, using "1" as fragments: 1st: 11 2nd: 1111 3rd: 11111111 4th: 1111111111111111 5th: 11111111111111111111111111111111 6th: 111111111111111111111111111111111111111111111111111111111111111

What are the features of Watson and Crick's structure of DNA?

It is a polymer of nucleotides. The polymer monomers are connected via deoxyribose sugars and phosphates. The nucleotide polymer strand is asymmetric because one end (5') has a phosphate while the other end (3') has a hydroxyl (OH). Complementary bases pair in the center through hydrogen bonds. So if the sequence of one polymer is known the other sequence can be derived. A pairs with T and G pairs with C. The two strands of DNA are connected in opposite orientation - they are antiparallel. DNA base sequence is a code for proteins.

In a DNA profile, why does one use a crime lab technician's DNA?

It serves as a control for the laboratory methods.

Why is it an advantage that many restriction-enzyme recognition sites read the same on either strand of the DNA molecule when read from the 59 end of the DNA? Remember: Restriction enzymes bind DNA and move along the molecule, looking for a recognition site.

Many recognition sites can be recognized by restriction enzymes on either strand of a double helix DNA molecule when read from the 5' direction. This property makes it possible for the restriction enzyme to identify these types of recognition sites from either direction. This makes it possible for the restriction enzyme to run along the DNA in either direction, and still preform the task of breaking a DNA molecule in two at the correct location.

Minisatellite

Nucleotide sequences 14 to 100 base-pairs long, organized into clusters of varying lengths, on many different chromosomes; used in the construction of DNA fingerprints.

The cloning methods outlined in this chapter allow researchers to generate many copies of a gene they wish to study through the use of restriction enzymes, vectors, bacterial host cells, the creation of genetic libraries, and PCR. Once useful quantities of a disease-causing gene are available by cloning, what kinds of questions do you think should be asked about the gene?

Once a gene is identified through the many types of methods and techniques, statistical questions should be asked about it. This way models can be built around it, and predictions can be made that will help manage the expression of it. Statistical questions about a disease-causing gene should take into account aspects like its prevalence in a population, its inheritance pattern, its penetrance, and its frequency in a population. Biochemical questions should be taken into account as well. What other genes does the gene in question interact with? What are its mechanisms for expression? Next, we should ask questions about its treatability, how to manage it? Is gene-therapy an option, and if so, is it ethical? Can the disease be treated in a fetus? There are many other factors that come into play that are not discussed here. When thinking about how to answer this question, remember to take into account all aspects of a disease. Try to imagine an individual's experience when coping with a genetic disease, the family too.

Parents undergoing IVF would not want to have an embryo with a genetic mutation implanted. What procedure would these parents ask the clinic to perform on the embryo?

PGD

Transgenic

Refers to the transfer of genes between species by recombinant DNA technology; transgenic organisms have received such a gene.

How was it established that induced pluripotent stem (iPS) cells derived from adult skin cells are really pluripotent?

Researchers discovered that adult stem cells have the capability to differentiate at the level of pluripotent stem cells. They discovered this by conducting an experiment using mice. They took cells from a mouse and turned them into induced pluripotent stem cells (iPS cells). From these iPS cells, the researchers were able to produce fertile adult mice. This proved that induced pluripotent stem cells had the capability of differentiating into any type of cell.

Vectors

Self-replicating DNA molecules that are used to transfer foreign DNA segments between host cells.

Short tandem repeat (STR)

Short nucleotide sequences 2 to 9 base-pairs long found throughout the genome that are organized into clusters of varying lengths; used in the construction of DNA profiles.

What was the first cloned dog called?

Snuppy

Why is it not possible to biopharm all needed proteins in bacteria?

Some proteins require extra help to fold. Molecular chaperones, the helper proteins, are only found in eukaryotes.

Adult stem cells

Stem cells recovered from bone marrow and other organs of adults. These cells can differentiate to form a limited number of adult cells and are called multipotent cells.

What is the purpose of technology?

Technology is the application of scientific knowledge for the betterment of society.

In a mass grave, how can you determine which bones belong to the children of a set of parents?

The DNA profiles of the children will have bands found in the DNA profile of one or both parents.

Name three of the many applications for Southern blotting.

The Southern blot is a technique used for analyzing cloned DNA. It is used to compare DNA sequences using probes that bind to known sequences. It can be used to analyze the differences in normal alleles versus mutant alleles. Southern blotting can be used to identify similar genes in different species of organisms. Southern blotting can also be used to study gene evolution, by identifying sequences present in genes and comparing their differences.

Dolly made headlines in 1997 when her birth was revealed. Why was cloning Dolly so important, considering that cattle and sheep were already being cloned through embryo splitting?

The birth of Dolly was a shock to the scientific community because she was the first mammal to be cloned using nuclear transfer from an adult somatic cell, which proved that the notion of differentiation is reversible. Prior to Dolly's birth, it was theorized that reversible differentiation was not possible by a cell. In other words, once a cell decided what type of cell it would become, it could not go back and turn into a different type of cell after it had developed and matured.

You are a neuroscientist working on a particular neuro-degenerative disease and discover that the disease is caused by the expression of a mutant gene in the brain. A colleague suggests that you use this new information to develop a transgenic mouse model of the disease. How might you do this? How would you use the animal to study the disease or look for new treatments?

The goal is to create a transgenic model mouse for a neurodegenerative disease caused by a mutant gene. First we would need to isolate the gene and clone it into a vector. Then we would have to inject the vector into the nucleus of a fertilized mouse egg. From there, the gene should be taken up into the mouse's genome and expressed throughout all the cells of the mature mouse. Using these transgenic model mice, we could draw linkages between brain structures and behavioral patterns. We could also screen drugs that could possibly manage or cure the disease in the mice and identify which ones are effective. Furthermore, we can identify molecular mechanisms that participate in the early stages of the disease. All of these things we could not ethically do with humans.

DNA profile

The pattern of STR allele frequencies used to identify individuals.

Why is PCR so revolutionary? Describe two applications of PCR.

The polymerase chain reaction is important because it increased the capacity of cloning quantity and speed, and in many cases, replaced host cell cloning. It revolutionized the technique of cloning, opening up more applications for it. For instance, the cloning of DNA can now be done with samples that previously were not sufficient in size or environment to clone. The application of PCR has made it possible to clone DNA from extinct animals. Another application is its use in clinical diagnosis. Clinical diagnosis can be made by analyzing a person's genetic makeup, which is made possible in little effort by PCR. Taking a sample and amplifying it from a patient can be as easy as a cheek swab and a PCR machine.

Multipotent

The restricted ability of a stem cell to form only one or a few different cell types.

Why is genetic screening done?

To catch babies with a genetic disease so they can be treated.

enzyme replacement therapy

Treatment of a genetic disorder by providing a missing enzyme encoded by the mutant allele responsible for the disorder.

True or False: If a suspect's DNA profile has bands that are not present in the crime scene sample (that does not match the victim's profile), then that person is no longer a suspect especially if another suspect's profile matches.

True

A new gene that causes an inherited form of retinal degeneration has been cloned from mice. This disease leads to blindness in affected individuals. As a researcher in a human vision laboratory, you want to see if a similar gene exists in humans (genes with similar sequences usually mean similar functions). With your knowledge of the Southern blot procedure, how would you go about doing this? Start with the isolation of human DNA.

We would like to analyze a gene that causes retinal degeneration in mice and determine if humans have a similar gene. To do this, we need a Southern blot of human DNA where we think we would find this gene and a Southern blot of the cloned mouse gene. We would need to use a probe in the Southern blot procedure that will bind to the mouse gene sequence that causes retinal degeneration. We already have an isolated form of the mouse gene in question. So first we need to obtain a DNA sample from the human and we can obtain one via cheek swab. With our samples of mouse and human DNA, we next run a polymerase chain reaction (PCR) with mouse and human DNA to amplify the amounts of copies we need, taking all the necessary steps to prepare the sample with restriction-enzymes. Now that we have many copies of DNA to work with, we run the sample DNAs through gel-electrophoresis, which separates the DNA into size. Next, we separate the double strands of DNA from each other in the gel. Now, we run a Southern blot procedure on the size separated DNA, which will transfer the DNA from the gel to DNA-binding filter paper. After the DNA has been transferred to the filter paper, we expose it to our probe, which will bind to the DNA (from the human and mouse) at known sequences associated with the mouse disease. We then wash away excess probe and expose it to X-ray films, which will produce a blotting pattern consistent with areas at which the probe attaches to DNA. Now we have a films with mouse and human DNA that will possibly expose similar sequences between them. This film will help us determine if there are sequences in human DNA that are associated with the gene in mice that causes retinal degeneration.

A base change (A to T) is the mutational event that created the mutant sickle cell anemia allele of beta globin. This mutation destroys an MstII restriction site normally present in the beta globin gene. This difference between the normal allele and the mutant allele can be detected with Southern blotting. Using a labeled beta globin gene as a probe, what differences would you expect to see for a Southern blot of the normal beta globin gene and the mutant sickle cell gene?

When a probe for the normal sequence is used, a Southern blot without a visible fragment for that sequence will be produced, because a base change in the gene sequence that produces sickle cell anemia has occurred. In this case, the probe will not bind to a sequence that is not identical to the normal sequence of nucleotides for that gene. So, because there is a base chance, the sequence is altered, and the probe is not able to bind to the sickle cell sequence. Furthermore, since the base chance for sickle cell anemia is at a restriction site, many large fragments will be produced. These large fragments are not normally found in the sample because the restriction enzyme is not able to identify the restriction site and cleave the DNA into smaller pieces.

You are given the task of preparing a cloned genomic library from a human tissue culture cell line. What type of vector would you select for this library and why?

Yeast artificial chromosome (YAC) vectors can be used to clone a human's entire genome because YAC vectors are derived from eukaryotic yeast cells that have telomeres and centromeres, which are able to hold long lengths of sequences. Yeast artificial chromosomes (YAC) are cloning vectors that are able to hold up to one million base pairs each. Since the human genome is so large, YAC vectors are suitable for cloning the human genome. A human genomic library can be created with about 30,000 YAC vectors, which is more efficient than using other types of vectors based on amount needed.

Answer the following questions with a yes or a no, and support each view with a coherent reason. a. Should very short children receive recombinant human growth hormone (HGH) to reach average height? b. Should genetically modified foods be labeled in grocery stores? c. Is it acceptable for physicians to treat children of normal height with HGH to make them taller if they and their parents want this?

a) The ethical use of human growth hormone (HGH) on short children is a matter of opinion. While considering its use, we must take into account for certain factors like safety, effectiveness, and physiological impact. How safe is the use of human growth hormone on a developing child? What are its long-term effects? Is it worth the risk of altering the biochemistry of a growing child to meet a social norm? We should also consider stigmas a person might face throughout life if he were to develop short. Would it be advantageous for the child to be taller, and is it worth the biological risk? We should consider these and other questions, along with scientific evidence when forming an opinion. b) Genetically modified foods are in wide use today. The labeling of genetically modified foods is not regulated, and some think it should be. Forming an opinion on this matter is subject to debate, and should consider many aspects. In forming an opinion we should consider the type of modifications, and their biological risks. What types of risks are there? Are there risks posed to the consumer and/or ecological sites in which the plants are grown? How well do we understand genetically modified foods? Is it just a misunderstanding of the science? We should consider these, and other questions, along with scientific evidence when forming an opinion. c) The ethical use of human growth hormone on normal children is a matter of opinion. When considering its use, we must take into account factors like safety, effectiveness, and physiological impact. How safe is the use of human growth hormone on a developing child? What are its long-term effects? Is it worth the risk of altering the biochemistry of a growing child to meet or exceed a social norm? We should also consider stigmas a person might face throughout life if he were taller. Would it be advantageous for the child to be taller, and is it worth the biological risk? Why is it (on average) more acceptable to use biotechnology on a deficient child, rather than to use it to produce advantages? We should consider these and other questions, along with scientific evidence when forming an opinion.

You are serving on a jury in a murder case in a large city. The prosecutor has just stated that DNA fingerprint-ing shows that the suspect must have committed the crime. He says that for cost reasons, two STR probes were used instead of four, but that the results are just as accurate. The first probe detected a locus that has a population frequency of 1 in 100 (1%), and the second probe has a population frequency of 1 in 500 (0.2%). a. What is the combined frequency of these alleles in the population? b. Does this point to the suspect as the perpetrator of the crime? c. Is there any other evidence you would like from the DNA lab? d. Do you think that most people on juries understand basic probabilities, or do you think that DNA evidence can be used to mislead jurors into reaching false conclusions?

a) To find the combined frequency of these two alleles in the population, we should multiply them together. Multiplication is used in this case because we are trying to find the chance of both alleles occurring at the same time (as opposed to one or the other). The first allele's frequency in the population is one percent, or 1 out of 100; also written 1/100. The second allele's frequency in the population is 0.2%, or 1 out of 500; also written 1/500. We multiply these two frequencies together: 1/100 × 1/500 = 1/50,000. This means one out of every 50,000 individuals will have this combination of alleles. b) As of 2012, Los Angeles, California had a population well over 3 million persons. This murder case is taking place in a large city, and if it were as large as Los Angeles, then using two STR probes that create a frequency of 1/50,000 would not point to the suspect as the perpetrator because the chances are too high that another person with the same alleles may exist in the same city. In a city of 3 million people, there are about 60 people with the same allele frequency. c) The lab should provide more probes to decrease the allele combination frequency, which would narrow down the chances that there are multiple people with the same allele combination, and therefore committed the crime. d) The jurors that use DNA fingerprinting evidence should understand the basic principals of the technique. This question is biased on opinion, and should be discussed.

The steps in the polymerase chain reaction (PCR) are: a. breaking hydrogen bonds, annealing primers, and synthesized using DNA polymerase b. restriction cutting, annealing primers, and synthesizing using DNA polymerase c. ligating, restriction cutting, and transforming into bacteria d. DNA sequencing and restriction cutting e. transcription and translation

a. breaking hydrogen bonds, annealing primers, and synthesized using DNA polymerase

What is meant by the term recombinant DNA? a. DNA from bacteria and viruses b. DNA from different sources that normally are not found together c. DNA from restriction-enzyme digestions d. DNA that can make RNA and proteins e. none of these

b. DNA from different sources that normally are not found together

A cloned library of an entire genome contains a. the expressed genes in an organism b. all the DNA sequences of an organism c. only a representative selection of genes d. a large number of alleles of each gene e. none of these

b. all the DNA sequences of an organism

Nuclear transfer to clone cattle is done by which of the following techniques? a. An 8-cell embryo is divided into two 4-cell embryos and implanted into a surrogate mother. b. A 16-cell embryo is divided into 16 separate cells, and those cells are allowed to form new 16-cell embryos that are implanted directly into surrogate mothers .c. A 2-cell embryo is divided into two separate cells and implanted into a surrogate mother. d. A 16-cell embryo is divided into 16 separate cells and fused with enucleated eggs. The fused eggs are then implanted into surrogate mothers. e. None of these.

d. A 16-cell embryo is divided into 16 separate cells and fused with enucleated eggs. The fused eggs are then implanted into surrogate mothers.

What was the first recombinant biopharming molecule?

insulin

Which DNA is used to make human proteins or copies of human genes in bacteria?

plasmid

pluripotent

the ability of a stem cell to form any fetal or adult cell type.

7. The following DNA sequence contains a six-base sequence that is a recognition and cutting site for a restriction enzyme. What is this sequence? Which enzyme will cut this sequence? (See Figure 13.5 for help.) 5' CCGAGGAAGCTTAC 3' 3' GGCTCCTTCGAATG 5'

the restriction enzyme that would work on the given piece of DNA is derived from the bacterium Haemophilus influenzae. The enzyme is known as HindIII and cuts between the following nucleotides. The original double strand of DNA has the sequence of: 5′ CCGAGGAAGCTTAC 3′ 3′ GGCTCCTTCGAATG 5′ The red nucleotides denote the recognition site for HindIII. HindIII cuts at the "/" in the following recognition sequence. A/AGCTT TTCGA/A The recognition site is in red text, and the backslash is where the enzyme cuts each nucleotide of the double strand. 5′ CCGAGGA/AGCTTAC 3′ 3′ GGCTCCTTCGA/ATG 5′

Embryonic stem cells and iPS cells are pluripotent meaning

they have the capacity to become any specific type of cell of the body because they have not yet differentiated.


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