215 #3

The 'F' DNA marker is closely linked to the B,b locus. The father is healthy, the mother has lung cancer. (A) What is the marker genotype of their four children? (B) Which two children are at the most risk for lung cancer? (C) Why are they at the most risk?

(A) 1 and 4 are ff, 2 and 3 are Ff (B) 1 and 4 (C) Children 1 and 4 most likely inherited the 'b' allele from their daddy

If the gene cloner can put 100 colonies on a plate, their genomic library of turtle genes (extracted from teenage mutant turtles) would need to have how many plates to be big enough to clone a mutant gene for Ninja traits.

100 - 500 (The number varies, depending on the genome size of the organism you are making the library for. A good guess for mutant turtles would be 100 - 500.)

Which place(s) can DNA pol llI add?

2

Refer to problem 2A on your problem set. Match plates 1, 2, 3 and 4 with the expected observations. (Match the outcome with the plate number)

2, 3, 4, 1 (2) will be a continual lawn of bacteria where colonies grew together (1) will contain no bacteria growth

Which bacteria was transformed?

A and B

DNA can be successfully extracted from:

All of the Above (A Silk Worm BT Corn Soil Bacteria) (Any living organism, or one that was alive at one time, contains DNA. Therefore, DNA can be extracted from all of the above including a silk worm, Bt susceptible corn, and soil bacteria.)

Function of this RNA/protein complex molecule is to

All of the above -Support molecules that are Forming hydrogen bonds -Form bonds between amino acids -Break bonds between amino acids and tRNAs

Why Is there no band In the lane 3 of the Western blot?

Antibody is specific for Bt which the RR does not have

If the gray DNA (A) is going to be used in gene cloning and is from a cow, the gene cloner must use a plasmid (B) that originated from a...

Bacteria (Plasmids are naturally found in bacteria and are used as vectors for moving genes/pieces of DNA into bacteria, where they will be copied.)

Best restriction enzyme to use in cloning Rhodopsin gene.

BlueII (The restriction enzyme should cut in the region that contains the marker gene so scientists can determine which bacteria have received a recombinant plasmid.)

Genetic engineering has what advantages over traditional plant breeding?

Both of the above. (A single gene can be transferred rather that many genes. It can cross the 'sexual barrier'. Traits can be transferred from any organism.) (Genetic engineering physically removes the DNA from one organism and transfers it to another removing the need for sexual mating and overcoming the 'sexual barrier' between species. It is also more specific. A single trait can be transferred rather than half of the genes present in the organism being mated.)

Which of these would be the first step in engineering a catfish that glows green because it has the transgene to encode jellyfish GFP?

Extract DNA from a jellyfish and identify the gene of interest (The first step is to obtain the DNA that will code for the desired trait)

The E. coli phytase enzyme is lane A. The other lanes are samples from different parts of the transgenic pigs. Which lane would be the stomach content?

G

The reason why a DNA sample that has a gene of interest is cut with a restriction enzyme in order to clone the gene is...

Genes are connected to other genes on the DNA molecule that makes up a chromosome so they need to be cut apart to clone a single gene. (Genes are a part of a chromosome that may contain thousands of genes and be millions of nucleotide pairs long.)

Plant breeding is a necessary step in genetic engineering because:

Genetic engineering does not work well on all elite lines. (Elite lines often cannot be efficiently transformed and cloned in tissue culture. It is more efficient to transform only a few lines and breed the transgene into other lines.)

This type of library contains promoters.

Genomic library (The starting material for a genomic library is DNA, while the starting material for a cDNA library is mRNA.)

During the genetic engineering process:

Living plant cells are changed genetically from the addition of new DNA. (Genetic engineering changes the genetic makeup of something that is already alive by the addition of foreign DNA.)

What is the amino acid represented by the star in the picture?

Met

Which plasmid should be used to clone a gene from corn if the gene in the corn DNA sample is cut into one fragment by the restriction enzyme PstI?

Neither would work.

An anticodon on a carnation tRNA is:3' - AUU - 5'This is the amino acid which it picks up in a Zebra Fish.

None -ile (isoleucine) -leu (leucine) -met (methionine)

What is the final product of transcription?

RNA

Enzyme that reads DNA and makes RNA.

RNA polymerase

Lane 2 is Whole eyeball proteins, lane 3 is rod cell proteins. The reason the protein bands look different is because...

Rod cells don't have all the mRNAs that the whole eyeball contains

To make a cDNA library to find the rhodopsin gene that causes color blindness in people, which cells should be the starting material?

Rod cells from a color blind boy (All DNA is found in every cell, and proteins are expressed in specific cells based on their function. mRNA codes for proteins.)

Sticky ends of DNA are...

Two ends of a double stranded DNA molecule that have overhanging nucleotides on one strand so that they can stick to complementary pieces by forming hydrogen bonds. (Ends of two DNA molecules will stick together if they have overhanding, complementary sequences.)

In the procedure shown below, the genetic engineer ....

a. will inject hundreds of copies of the transgene into this fertilized egg.

What can you do to make sure you have functional oncogenes?

all of the above (do not smoke, wear sunscreen limit exposure to X-rays) (Phenotype is affected by genotype and environment)

What forces that we might encounter in the environment cause mutations?

all of the above are mutagens (UV light, Ionizing radiation certain chemicals)

BRCA-1 is an anti-oncogene that helps repair damaged DNA in breast cells. Why would a person have a test done to determine what BRCA-1 alleles they have in their cells.

all of the above would be reasons to have an anti-oncogene test such as the test for BRCA-1 (Genetic information can tell us about inheritance and risk of cancer. Anti-oncogenes are cancer-fighting genes.)

BRCA-1 is an anti-oncogene that helps repair damaged DNA in breast cells. Why would a person have a test done to determine what BRCA-1 alleles they have in their cells.

all of the above would be reasons to have an anti-oncogene test such as the test for BRCA-1 (they are a member of a family in which breast cancer occurs in successive generations. the test results can help them determine if they are at a higher risk of getting breast cancer. the results of the test can guide their choices in determining how often they might have testing for early signs of breast cancer.)

The RNA polymerase is made of...

amino acids.

Using this plasmid, the color of non-recombinant colonies on a selective media plate:

blue

A mechanism used to minimize changes in DNA sequence in an organism is...

both of the above (Proof reading by DNA polymerase III to minimized replication mistakes. Repair of damaged DNA by repair enzymes.)

A mechanism used to minimize changes in DNA sequence in an organism is...

both of the above (Proof reading by DNA polymerase III to minimized replication mistakes. Repair of damaged DNA by repair enzymes.) (DNA Polymerase III reads from 3' to 5' and builds from 5' to 3')

When the bacteria in A is transformed...

it is given a few more genes than it already had. (It acquires just a few more genes; those located on the plasmid it was transformed with (ie antibiotic resistance and lacZ).)

A mutation occurs in a gene and the cell with this mutant allele no longer controls it's cell cycle. Uncontrolled cell division results. The gene that was mutated is a...

oncogene

A mutation occurs in a gene and the cell with this mutant allele no longer controls it's cell cycle. Uncontrolled cell division results. The gene that was mutated is a...

oncogene (Oncogenes control the cell cycle. Anti-oncogenes code for cancer-fighting proteins.)

How much time would it take to go from A to B?

overnight (Letting E. coli incubate overnight allows the cells enough time to grow and multiply.)

A cDNA clone can be used to sequence a protein . A genomic clone can be used to sequence a promoter .

protein promoter

In order to design the gene that will be expressed in the desired part of the cow, the genetic engineers should....

use a casein promoter and the strawberry flavor coding region. (The promoter will bind to the DNA of the organism you are genetically engineering. Therefore, the promoter should share DNA with that organism. The coding region should come from the organism with the trait of interest, since the genetic material from the coding region will determine the traits expressed in your genetically engineered organism.)

An oncogene in a cell mutates and the cell becomes cancerous. The person in which this happens....

. will be able to fight off these cancer cells by the action of anti-oncogenes.

There are 6 map units between the 'F' marker and the B,b Locus. What is the chance that 2 and 3 will get cancer (are bb)? HINT: Take into account what you know about their genotype.

.03/.5 = 6%

The Rhodopsin protein is 700 amino acids long. How many nucleotides are there in the Rhodopsin gene coding region. (minimum number.)

2100 + 3 = 2103, if you count the Stop codon

You want to make a copy of the DNA segment shown from the Primer 1 to the Primer 2 binding regions. What should be the sequence of Primer 1?

5'CTCCTAAA3' (The primer will build nucleotides that are complimentary to the DNA strand, reading from the 3' to the 5' end.)

Lane 3 is Rod cell proteins. Which protein is found in the largest amount?

A

If you want to make a library that contains a gene you are interested in, you should...

A and B above are true. (Start with DNA from the organism you know has this gene. Plan to make a library of thousands and thousands of bacteria colonies that each has cloned DNA from the organism that has this gene.) (Because we cannot separate the gene of interest away from all the other DNA sequences at the early stages of gene cloning, the steps described in #a and #b are necessary.)

Once the pig geneticists had lots of copies of the transgene... what kind of lab was the next place they did their work in order to create Enviropig?

A lab with a microscocpe, very fine needles and a suction tube that was microscopic

Which step in the gene cloning process is done in a living cell?

A recombinant plasmid is copied many times. (Living bacteria cells make copies of recombinant plasmids that contain genes from another organism. This is how entire genes are cloned.)

During genetic engineering of a plant a new gene is inserted into:

A single cell

In genetic engineering, gene cloning is used to make a copy of _____________?

A single desirable gene controlling the desired trait. (When DNA is extracted from an organism, all its DNA is extracted. Gene Cloning is used to locate and make copies of the one gene of interest.)

In order for genetic engineering to work, DNA must be:

A stable molecule so it can be removed from the cells of one organism, transferred to a new cell, and maintain it's function. (The DNA molecule is removed from the cell and handled many times during genetic engineering. It must be stable.)

From which of the following organisms could genes that are used to genetically engineer crop plants originate?

All of the above (Bacteria Humans Another plant)

Why do field mice have a lower risk of skin cancer than frogs that live in the same part of Nebraska?

All of the above will contribute to mice having a lower risk of skin cancer than frogs. (Increased sunlight exposure increases the risk of a mutation occurring in the oncogenes.)

Below are three occupations or life styles. A person with which occupation/life style would have the most spontaneous mutations cause by mistakes in DNA replication?

All of the above would be about the same. (coal miner who works the day shift life guard who works on a beach in Hawaii. A bald Nebraska farmer who enjoys topless field work.)

Plasmids are useful cloning vectors because they...

All of the above. (are small, mobile DNA molecules. can be combined with other DNA molecules and will be replicated in bacteria cells. have genes that serve as selectable markers because they change easy to detect traits in bacteria cells.)

If you left the X-gal out of the media plates below...

All the colonies would be white (Without X-gal present, no color will be formed. There would be no substrate for the lac Z gene to act upon and therefore, no way of distinguishing bacteria transformed with recombinant plasmids and those transformed with non-recombinant plasmids.)

What would you be able to obtain from a cDNA clone, but not from a genomic clone of the same gene?

Amino acid sequence of the protein encoded by the gene (Since a cDNA clone is made from mRNA, the intron sequences will have already been spliced out. As a result, the bacteria cell which is transformed with this cDNA clone will be able to make the correct amino acid sequence for the protein.)

These are the type(s) of gene libraries that can be screened with the method shown.

Answers b, c and d are all correct. -cDNA library of a eukaryote -Genomic library of a prokaryote -cDNA library of a prokayote

To make a genomic library to find the rhodopsin gene, which cells should be the starting material?

Any cells above will work (Blood cells from the mom of a color blind boy Eyeball cells from pigs. Rod cells from a color blind boy)

Place these historical events in biology/genetics the correct order, earliest first. A. Scientist invented PCR B. Scientists showed DNA could transform bacteria C. Scientists learned how to extract nucleic acids from cells, but not sure what the stuff was D. Based on the complexity of the molecules, biologists believed the genetic material was protein and not DNA E. Scientists discovered the structure of DNA

C D B E A

Blue colored corn is the "normal" color of corn. Yellow corn resulted from a mutation in the gene causing blue color protein to be too short and not function at all.This mutation would be occuring in what part of the gene?

Coding region exon (Genetic material that enters the cytoplasm and undergoes transformation will affect gene expression.)

Assume you are trying to obtain a genomic clone of the red fluorescent protein (RFP) gene from coral. What item in the list of lab materials (in the Problem Set) would you need to use as your starting material for a library that will contain the RFP gene clone?

DNA from coral cells

Three living things that are targets for genetic engineering: 1.) Dogs 2.) Rice Plants 3.) E. coli Bacteria 4.) Catfish Successful genetic engineering requires gene insertion and then the propagation of a new organism from the transformed cell. The best Ranking of these organisms from the easiest to the most difficult is....

E. coli then Rice then Catfish then Dogs (Think about: 1) The appropriate technique for transforming each kind of organism. 2) The gestation period of that organism. 3) The ability to access the type of cell that will be transformed for each organism.)

The E. coli phytase enzyme is lane A. The other lanes are samples from different parts of the trangenic pigs. Which lane would be the stomach lining cells?

F

A person has a somatic mutation and they get skin cancer. Will they be able to pass this skin cancer mutation on to their offspring.

No (Somatic cells are all non-reproductive cells in an organism. Gametic cells are the sex cells and will be inherited.)

DNA sequence of the RNA-like strand of a gene coding region for normal enzyme (before intron sequence is encountered).5'CCCATGTTTAAACCCCGC............3'If the first G on the 5' end of this sequence is changes to a C in a mutant allele, the result would be...

Normal: Met Phe Lys Pro Arg ......Mutant: not sure since a different start codon would be found on the mRNA (Your amino acid chart will tell you which amino acids are coded from different combinations of three nucleotides. Mutations in the nucleotides may change an individual amino acid or many amino acids.)

If the Holding team of maize geneticists wanted to determine the difference in the length and abundance between the mRNA than encode the 22kD-s protein and the 19KD-s protein they should use which technique(s)?

Northern blot RT-PCR

What is the difference between an oncogene and an antioncogene?

Oncogene encode proteins needed to control the cell cycle while antioncogenes encode cancer fighting or DNA repair proteins.

Fill in the blanks. Gel X is the ____ gel of the transgene, gel Y is the ______ gel of the transgene. The Western Blot would look similar to _____

PCR, RT-PCR, gel Y

Genectic engineering works by:

Physically removing the DNA that controls a trait and transferring it to another organism.

GFP Zebra fish can be a bioreactor, assume these zebra fish have the transgene shown below

Positive, On

Your biotechnology company has patent rights on the genes below. Your gene designer needs to create a transgene that can be used to transform cows that make milk that could protect the milk drinker from west nile virus. The transgene should have....

Promoter of E and the coding region of D. (The promoter should have genetic information that is compatible with the organism of interest and the coding region should contain genetic information associated with the desired trait.)

Your company has patent rights on the genes below. In order to make onion flavored transgenic apples, they should design a transgene that combines...

Promoter of H and coding region of G (The promoter should have genetic information that is compatible with the organism of interest and the coding region should contain genetic information associated with the desired trait.)

Ligase enzyme ...

Seals sugar phosphate backbone bonds on DNA molecules and is thus a useful enzyme in gene cloning. (Ligase can function in a test tube to stabilize a recombinant DNA molecule by sealing the sugar phosphate backbone bonds once two molecules come together.)

Plasmids are...

Small circular DNA molecules that usually contain two or a few genes. (Plasmids are cloning vectors because they are small DNA molecules and thus have the mobility to be transferred to bacteria cells.)

These are differences between using gene therapy vs. genetic engineering in animals.

Some cells vs. all cells NOT vs. inherited Somatic cells vs. egg or embryo

A palindrome would have a sequence AAAGCTTT and is found in this part of the Rhodopsin gene

Termination sequence

Modification of a gene (gene design) is done where?

Test tube (Gene modification is done to extracted DNA, which would be stored in a test tube. Special enzymes are added to the test tube to cut apart the gene regions and replace them with desirable regions.)

Genetic engineering is:

The addition of foreign DNA to an organism's genome

Assume that a person inherits a mutant allele of the anti-oncogene p53. As a result they might experience which of the following?

The will have a higher risk of cancer or certain kinds of cancer in their lifetime. (Anti-oncogenes are cancer-fighting genes.)

RNA is made 5' to 3'

True

What mutation would have the biggest potential impact on your body?

a mutation in a gene that encodes a cell cycle regulation protein. This causes the cell to no longer go through and orderly cell cycle. (Cells replicate and replace dead cells. Therefore, a mutation affecting the replication process would have a high impact on organismic function.)

P53 is a transcription factor (regulatory) protein that turns on cancer cell fighting genes. What kind of gene is p53, besides a regulatory gene?

anti oncogene

Arrange these steps in the proper order to describe the function of the araC and the GFP genes on this cloning vector. _____a. araC transcription factor protein binds to GFP promoter _____b. araC gene is transcribed _____c. arabinose sugar binds to the araC transcription factor protein _____d. araC protein falls off the GFP promoter _____e. araC mRNA is translated _____f. GFP is transcribed, GFP mRNA is translated, GFP glows green inside the E. coli cell

b e a c d f

What type of animal transformation is most likely to give chimeric animals (animals with some genetically engineered cells and some that are not)

blastocyst embryonic cell transfection

What would be the next step in gene cloning after completing the step shown?

combine the two different tubes (The next step would be to combine the two tubes; plasmid and DNA of interest (which the library is being made from), both of which have been cut with the same restriction enzyme.)

Translation takes place in this part of a genetically engineered carnation flower cell

cytoplasm

A cDNA clone includes which of the following (select all of the ones that apply)?

exons

What happens with families that pass on mutant p53 genes?

higher risk of cancer

The transformation method used to produce the organism show in the diagram below would be....

injection with a fine needle into a fertilized pig egg.(Pronuclear injection is a technique used to introduce genetic material into animal embryos. Plants and animals differ, as cell of a plant can be transformed, while in animals, embryos must be targeted for transformation.)

Making a genetically engineered crop from beginning to end...

is difficult and may take many years. (The genetic engineering process is very time consuming, tedious and inefficient. Many steps still involve 'hit and miss' science that requires many tries before success is achieved.)

Which plasmid should be used to clone a gene from corn if the gene in the corn DNA sample is cut into one fragment by the restriction enzyme KPNII?

pKSU (Both plasmids have one site where KpnII will cut but an insert will inactive a gene in pKSU but not in pUNL.)

Assume the maps are for two plasmids that can be used in gene cloning. The maps show the restriction enzyme cutting sites on each plasmid and the locations of genes that will confer resistance to amphicillin or encode an enzyme that will catalyze a color if the proper substrates are added to the media. Which plasmid should be used to clone a gene from corn if the gene in the corn DNA sample is cut into one fragment by the restriction enzyme HindIII?

pUNL (pUNL has one site the HindIII will cut and an insert in that site will inactivate the red color gene, allowing for white colony selection.)

UV light hits DNA in a skin cell and causes a Thymine dimer. As a result...

repair endonucleases cut out the thymine dimer and replace this with the right nucleotides.

In the process of genetic engineering, the gene design step...

results in desirable expression of the gene once inside a cell. (By modifying the gene construct, genetic engineers can design a gene that will act in a desired way once inside a plant. This ability gives them countless options to try.)

How are tRNAs made?

transcription of tRNA genes in the nucleus

The picture below is showing....

transformation of an animal (Pronuclear injection introduces new genetic material into embryos)

The E. coli phytase enzyme is lane A. The other lanes are samples from different parts of the transgenic pigs. Which phytase enzyme is longer?

transgenic version of E. colli