AP Bio Chapter 20 Biotechnology
2 advantages of using yeast cells over bacteria
1. YAC's: can hold a longer segment of DNA or the entire gene 2. Euk cell so it already contains the necessary elements for Euk gene expression 3. Has a plasmid and replicates quickly
The human genome is thought to contain about how many genes? (Concept 20.4E-Book) [Hint] 1,000-2,000 4,000-6,000 13,000-15,000 25,000-30,000 100,000-110,000
25,000-30,000
What characteristics would you want in an enzyme for use in a recombinant plasmid?
A staggard cut that would create sticky ends on both genes not in the middle of them or the origin of replication
Once an organism's genome sequence has been determined, how do scientists generally start identifying all the genes within the genome? (Concept 20.4E-Book) [Hint] Analyze the sequence using software that scans the sequence for telltale sequence elements such as promoters, transcription start and stop sites, and so on. Hybridize the DNA with probes specific for known genes from other organisms. Use DNA microarrays to examine the expression of huge numbers of sequences from the genome under different conditions. Individually mutate all of the nucleotides within the genome, and determine which mutations cause a detectable phenotype.
Analyze the sequence using software that scans the sequence for telltale sequence elements such as promoters, transcription start and stop sites and so on.
Besides the gene of interest, what other gene would you put in a recombinant plasmid?
Antibiotic resistance gene
To create recombinant DNA with long-term stability, it is necessary to have which of the following in the test tube? (Concept 20.1E-Book) [Hint] hydrogen bonding DNA ligase reverse transcriptase DNA polymerase heat-resistant DNA polymerase
DNA ligase
Which one of the following enzymes could seal a nick in one strand of a double-stranded DNA molecule by creating a sugar-phosphate bond between the adjacent, unjoined nucleotides? (Concept 20.1E-Book) [Hint] DNA ligase reverse transcriptase restriction enzyme terminator enzyme DNA polymerase
DNA ligase
Human nerve cells differ from human muscle cells because different sets of genes are expressed; in each type of cell, different genes are transcribed into mRNA and translated into protein. Which of the following techniques would be the most efficient way to identify the genes that these cells express? (Concept 20.4E-Book) [Hint] gel electrophoresis of DNA fragments DNA microarray assays isolating and analyzing all the proteins from each type of tissue PCR chromosome walking
DNA microarray assays
Advances in which of the following technologies was most helpful in the study of eukaryotic genomes? gene cloning DNA sequencing southern blotting transformation
DNA sequencing
Linkage mapping, physical mapping, and ________ each contributed to the genome mapping project by arranging fragments of DNA In order on increasingly smaller scales. DNA sequencing gene cloning electrophoresis PCR Southern Blot gene mapping hybridization
DNA sequencing
Briefly describe a method of isolating a plasmid?
Disrupt the cell membrane and then use gel electropherisis to isolate the plasmid
A scientist wishing to create an organism capable of breaking down several kinds of toxic waste combines genes from several species of bacteria to create a single "superbacterium." Which of the following would probably not be needed to do this? (Concept 20.1E-Book) [Hint] nucleic acid probes F factors plasmids restriction enzymes DNA ligase
F factors
What is the formula for writing the name of enzymes?
First letter of the genus+ first and second letter of the species + strain + roman numeral
Electrophoresis-->Southern Blotting-->Hybridization -->Autoradiography
Have to have all, there really isn't a point in doing only one or two of them.
If a portion of a gene's sequence is known, small, and a single stranded DNA called a probe can be made and labeled with a radioactive or fluorescent tag. DNA sequencing gene cloning electrophoresis PCR Southern Blot gene mapping hybridization
Hybridization
What is cDNA, and what is it used for?
It is DNA synthesized from mRNA by the enzyme reverse transcriptase that only contains exons. It can be used to clone eukaryotic genes in prokaryotes.
Autoradiography
It will turn dark on the x-ray film where the RFLP bands hybridized on it.
Amplifies small amounts of DNA rapidly by a heating/cooling cycle. DNA sequencing gene cloning electrophoresis PCR Southern Blot gene mapping hybridization
PCR
Each repeat of the cycle doubles the number of copies of DNA. DNA sequencing gene cloning electrophoresis PCR Southern Blot gene mapping hybridization
PCR
SSDNA primers are added along with DNA polymerase and nucleotides, which replicate DNA on each exposed segment as the solution is cooled. DNA sequencing gene cloning electrophoresis PCR Southern Blot gene mapping hybridization
PCR
RFLPs can be extracted onto a nitrocellulose transparency DNA sequencing gene cloning electrophoresis PCR Southern Blot gene mapping hybridization
Southern Blot
A molecular biologist used a retroviral vector to introduce a gene coding for a certain human enzyme into mouse cells. One cell line was isolated that was able to make the human enzyme, but it had lost the ability to express an endogenous, normally expressed gene in the process. What is the best explanation for these results? (Concept 20.5E-Book) [Hint] The virus caused the mouse cells to become diseased. The virus had transferred a gene from one mouse cell to another. The virus inserted the gene encoding the human enzyme into the normally expressed endogenous gene. The virus was too small to carry the entire gene. The enzyme acted as a nuclease enzyme, cutting up mouse DNA.
The virus inserted the gene encoding the human enzyme into the normally expressed endogenous gene.
DNA microarray assay
Use this to find out which genes are being expressed at what time
A genetic marker is _____. (Concept 20.2E-Book) [Hint] a place where a restriction enzyme cuts DNA a chart that traces the family history of a genetic trait a particular nucleotide sequence whose inheritance can be followed a radioactive probe used to find a gene an enzyme used to cut DNA
a particular nucleotide sequence whose inheritance can be followed
In recombinant methods, the term "vector" refers to _____. (Concept 20.1E-Book) [Hint] the enzyme that cuts DNA into restriction fragments the "sticky" ends of a DNA fragment an RFLP marker a plasmid or other agent used to transfer DNA into a living cell a DNA probe used to locate a particular gene
a plasmid or other agent used to transfer DNA into a living cell
Which of the following would be considered a transgenic organism? (Concept 20.5E-Book) [Hint] a bacterium that has been treated with a compound that affects the expression of many of its genes a human treated with insulin produced by E. coli bacteria a fern grown in cell culture from a single fern root cell a rat with rabbit hemoglobin genes all of the above
a rat with rabbit hemoglobin genes
What two enzymes are needed to produce recombinant DNA? (Concept 20.1E-Book) [Hint] a restriction enzyme and a topoisomerase a restriction enzyme and a ligase a restriction enzyme and a polymerase a polymerase and a ligase a polymerase and a topoisomerase
a restriction enzyme and a ligase
Southern blotting is _____. (Concept 20.2E-Book) [Hint] a method of DNA amplification a technique used to study RFLPs how bacteria take up DNA from the surrounding solution the insertion of DNA into a plant's chromosomes used to determine the product of a particular gene
a technique used to study RFLPs
Gene therapy involves _____. (Concept 20.5E-Book) [Hint] adding a functioning version of a defective gene to the cells of an individual allowing individuals to follow the natural progression of a genetic disorder, accompanied by psychological counseling, then drug treatment when the condition becomes life-threatening no serious ethical questions replacing organs affected with genetic disorders by transplants all of the above
adding a functioning version of a defective gene to the cells of an individual
The number of genes in an organism's genome is not a perfect indication of the organism's complexity because _____. (Concept 20.4E-Book) [Hint] alternative splicing can increase the number of polypeptides made from a single pre-mRNA post-translational modifications can increase the types of proteins produced by a single gene individual polypeptides can interact to form multiple types of protein complexes individual genes can be expressed in more or less complex ways in different organisms all of the above
all of the above
DNA synthesized using an RNA template is called _____. (Concept 20.1E-Book) [Hint] reverse transcriptase a restriction enzyme cDNA a palindrome a plasmid
cDNA
Bacteria use restriction enzymes to _____. synthesize DNA synthesize RNA synthesize protein destroy foreign protein destroy foreign DNA
destroy foreign DNA
In the polymerase chain reaction (PCR), the sequence of bases in the primers is important because it _____. (Concept 20.1E-Book) [Hint] determines which segment of the genome will be amplified always matches a stop codon always causes a silent mutation determines how many cycles of the reaction are needed to obtain a sufficient amount of amplified DNA determines the number of tandem repeats in a genome
determines which segment of the genome will be amplified
After digestion with restriction enzymes, the various resulting DNA fragments (negatively charged) are pulled through a gel matrix. DNA sequencing gene cloning electrophoresis PCR Southern Blot gene mapping hybridization
electropheresis
Results in RFLP's. DNA sequencing gene cloning electrophoresis PCR Southern Blot gene mapping hybridization
electropheresis
Longer fragments move more slowly, so all fragments are separated by size DNA sequencing gene cloning electrophoresis PCR Southern Blot gene mapping hybridization
electrophoresis
RFLPs can serve as genetic markers: the frequency with which two RFLP markers are inherited is a measure of the closeness of their two loci on a chromosome. DNA sequencing gene cloning electrophoresis PCR Southern Blot gene mapping hybridization
gene mapping
A nucleic acid probe is used to _____. (Concept 20.1E-Book) [Hint] clone genes produce a large amount of DNA from a tiny amount of DNA make exact copies of DNA sequences identify genes that have been inserted into bacterial plasmids or separated by electrophoresis make DNA from RNA
identify genes that have been inserted into bacterial plasmids or separated by electrophoresis
In genetic engineering, the highly active plasmid from Agrobacterium tumefaciens is used to _____. (Concept 20.5E-Book) [Hint] insert genes of interest into plant chromosomes cut DNA at a specific base sequence locate specific genes on animal chromosomes detect and correct mistakes in DNA replication do all of these
insert genes of interest into plant chromosomes
Which of the following is not a step of the Southern blotting procedure? (Concept 20.2E-Book) [Hint] hybridizing the DNA with a radioactive probe digesting the DNA with a restriction enzyme separating the DNA fragments using gel electrophoresis transferring the DNA to a blot linking the DNA with DNA ligase
linking the DNA with DNA ligase
Separating DNA fragments by gel electrophoresis is useful for all of the following except _____. (Concept 20.2E-Book) [Hint] identifying DNA fragments for RFLP analysis purifying specific DNA fragments distinguishing between different alleles of a gene identifying a plasmid or a virus by examining its restriction fragment pattern none of the above
none of the above
A molecular biologist has isolated a short segment of DNA that she wants to replicate in vitro. First she heats the DNA, which separates the two strands, and then she adds _____. (Concept 20.1E-Book) [Hint] nucleotides, primers, and polymerase ribosomes, nucleosomes, and messenger RNA nucleotides and ligase transfer RNA, matching amino acids, and messenger RNA ribosomes, matching amino acids, and primers
nucleotides, primers, and polymerase
Preparing a physical map of the genome involves _____. (Concept 20.3E-Book) [Hint] ordering markers such as RFLPs within the genome ordering markers such as simple sequence repeats within the genome preparing a collection of large, overlapping genomic DNA fragments, and ordering them relative to each other determining the nucleotide sequence of small genomic fragments all of the above
preparing a collection of large, overlapping genomic DNA fragments, and ordering them relative to each other
The dideoxyribonucleotide chain-termination method _____. (Concept 20.3E-Book) [Hint] produces a ladder of DNA fragments, with each individual band labeled with one of four different fluorescent tags can be used to sequence entire eukaryotic chromosomes in a single reaction is very slow, requiring several weeks to determine a sequence of about 200 nucleotides does not involve electrophoresis is difficult to automate and must be performed under close human supervision
produces a ladder of DNA fragments, with each individual band labeled with one of four different fluorescent tags
An enzyme that cuts DNA at a symmetrical sequence of bases is called a _____. (Concept 20.1E-Book) [Hint] reverse transcriptase restriction enzyme cDNA palindrome ligase
restriction enzyme
What is the source of the reverse transcriptase used in recombinant DNA technology? (Concept 20.1E-Book) [Hint] retroviruses plant cells cultured fungal cells cultured phage-infected mammalian cells either retroviruses or cultured phage-infected mammalian cells
retroviruses
Which arrangement of the following four enzymes represents the order in which they would be used in a typical gene-cloning experiment resulting in the insertion of a cDNA into a bacterial plasmid? Begin with the gene's mRNA transcript. (Concept 20.1E-Book) [Hint] restriction enzyme, reverse transcriptase, DNA polymerase, DNA ligase restriction enzyme, DNA ligase, reverse transcriptase, DNA polymerase reverse transcriptase, DNA polymerase, restriction enzyme, DNA ligase reverse transcriptase, DNA ligase, DNA polymerase, restriction enzyme reverse transcriptase, restriction enzyme, DNA polymerase, DNA ligase
reverse transcriptase, DNA polymerase, restriction enzyme, DNA ligase
In genetic engineering, "sticky ends" refers to _____. (Concept 20.1E-Book) [Hint] a technique for finding a gene of interest within a nucleus without destroying the cell the ability of plasmids to stick to a bacterial cell wall and thus be taken up into the bacterium short bits of single-stranded DNA left at the end of DNA molecules cut by restriction enzymes the site on mRNA that sticks to the DNA during transcription none of the above
short bits of single-stranded DNA left at the end of DNA molecules cut by restriction enzymes
DNA fingerprints are used to determine whether Sam could be the father of Becky's baby. Sam is not the father if _____ genetic fingerprint shows some bands not present in _____ genetic fingerprint. (Concept 20.5E-Book) [Hint] Sam's ... the baby's Becky's ... the baby's the baby's ... Sam's the baby's ... Becky's the baby's ... Sam's or Becky's
the baby's ... Sam's or Becky's
When a typical restriction enzyme cuts a DNA molecule, the cuts are staggered so that the DNA fragments have single-stranded ends. This is important in recombinant DNA work because _____. (Concept 20.1E-Book) [Hint] it allows a cell to recognize fragments produced by the enzyme the single-stranded ends serve as starting points for DNA replication the fragments will bond to other fragments with complementary single-stranded ends it enables researchers to use the fragments as introns only single-stranded DNA segments can code for proteins
the fragments will bond to other fragments with complementary single-stranded ends
Transgenic organisms can be scientifically or commercially useful only if _____. (Concept 20.5E-Book) [Hint] the inserted ("foreign") gene is drawn from the human genome the inserted ("foreign") gene is expressed in the host organism the host organism is a microorganism the vector is a plasmid all of the above conditions are met
the inserted ("foreign") gene is expressed in the host organism
DNA fingerprints used as evidence in a murder trial look something like supermarket bar codes. The pattern of bars in a DNA fingerprint shows _____. (Concept 20.5E-Book) [Hint] the order of bases in a particular gene the presence of various-sized fragments of DNA the presence of dominant or recessive alleles for particular traits the order of genes along particular chromosomes the exact location of a specific gene in a genomic library
the presence of various-sized fragments of DNA
RFLPs have been tremendously useful for genomic mapping studies because _____. (Concept 20.2E-Book) [Hint] they are found only in the coding sequences of genes they are found only in the promoter regions of genes they are found only in disease-causing genes they are not restricted to genes, and are abundantly scattered throughout the genome they are found only in expressed genes
they are not restricted to genes, and are abundantly scattered throughout the genome
In the polymerase chain reaction (PCR) technique, a heating phase and a cooling phase alternate. An original sample of DNA would have to pass through how many total rounds of heating and cooling before a sample is increased eight times in quantity? (Concept 20.1E-Book) [Hint] two three four six eight
three
Why would that gene be desirable?
to provide resistance to antibiotics so the cell can survive and replicate. Also, only transformed germs will grow on the antibiotic medium.
Because eukaryotic genes contain introns, they cannot be translated by bacteria, which lack RNA-splicing machinery. But if you want to engineer a bacterium to produce a eukaryotic protein, you can synthesize a gene without introns. A good way to do this is to _____. (Concept 20.1E-Book) [Hint] alter the bacteria so that they can splice RNA use a nucleic acid probe to find a gene without introns work backward from mRNA to make a version of the gene without introns use a phage to insert the desired gene into a bacterium use a restriction enzyme to remove introns from the gene
work backward from mRNA to make a version of the gene without introns
