Bio Honors Name:Date:Period:

Genetic Engineering Worksheet

1.The following segments of DNA have been obtained from three different individuals, A, B, and C respectively. The arrows indicate sequences where restriction enzymes would cut the DNA. Use the information in the diagram to determine fragment number and size per person and draw what the electrophoresis gel might look like if you were to run the DNA through this process.

2. Draw in bands to indicate the location of the DNA fragments that would be formed in a gel electrophoresis (from each segment above) on the gel below. Label the side of the gel with numbers to indicate the number of nucleotides in each band.(Bands with the same number of nucleotides should be shown at the same distance from the wells.) Also, indicate which side of the gel box would be positively charged and which would be negatively charged.

Use the following gel results to answer question #3:

3.Determine if paternity has been established for both Case 1 and Case 2:

  1. Case 1:
  2. Case 2:

4.Which of the following is the most logical sequence of steps shown for splicing foreign DNA into a plasmid and inserting the plasmid into a bacterium

  1. Transform E. coli cells
  2. Cleave (cut) by restriction enzyme
  3. Extract plasmid from bacterial cells
  4. Join plasmid DNA with foreign DNA by hydrogen bonds
  5. Seal with DNA ligase
    ______

5.Which of the following pieces of DNA could be cleaved (cut) by restriction enzymes to produce “sticky ends”?

  1. A AA G GGd. G A A T T C
    T TT C CC C T T A A G
  2. C A G C A G e. G C T T A C
    G T C G T C C G A A T G
  1. C CC T TT
    G GG A AA

6.Suppose a pharmaceutical company planned to clone the gene for human phophriobsyltransferase, an important enzyme. Which of the following steps would not be involved in this process?

  1. Joining the sticking ends with the enzyme DNA ligase
  2. Cutting human DNA with a restriction enzyme
  3. Cutting human DNA into individual nucleotides
  4. Transforming bacteria with recombinant plasmid/human DNA
  5. Joining human DNA with a vector such as a plasmid

Matching

7.Which of the terms at the right would be needed for the following two processes? Select all possible answers.

DNA Fingerprinting
______

Cloning a gene to produce a protein product

______

When circular plasmids are cut with restriction enzymes, different sized fragments of DNA are produced (NOTE: when a single cut is made to a circular plasmid, only 1 fragment is produced; while a single cut to a linear (straight) piece of DNA produces 2 fragments)

Use the following figure to answer questions 8-11. The plasmid pGEN101 shown was treated with various mixtures of restriction enzymes. The electrophoresis gel shows the results of each of those digestions.

8.Which lane (I to V) represents the fragments produced using Bam HI only? ______

9.Which lane represents the fragments produced using Eco RI only? ______

10.Which lane represents the fragments produced when the plasmid was cut with both Eco RI and Bam HI? ______

11.It is theoretically possible for a gene from any organism to function in any other organism. Why is this possible?

  1. All organisms have the same genetic code
  2. All organisms are made up of cells
  3. All organisms have similar nuclei
  4. All organisms have ribosomes
  5. All organisms have transfer RNA

Use the following diagram to answer questions 12-13.

12.The DNA fingerprint above represents 4 individuals. Which of the following statements is consistent with the results

  1. B is the child of A and C
  2. C is the child of A and B
  3. D is the child of B and C

13.Each single band in a banding pattern on a gel represents:

  1. One chromosome
  2. One piece of DNA
  3. Many pieces of DNA that are the same size
  4. Many chromosomes that are the same size
  5. Many pieces of DNA that are different sizes

14.The segment of DNA pictured below has been cut with specific enzymes which create fragments A, B, and C.

Which of the following gels produced by electrophoresis would represent the separation and identity of these fragments?

Use the following diagram to answer questions 15-16.

15. How many restriction sites were recognized by the enzyme that cut linear DNA that was run in lane 2? ______

16. Assuming that all fragments are visible, how many restriction sites were recognized by the enzyme that cut plasmid DNA in lane 3? ______

This last question is more difficult, but try it!

The following electrophoresis gel shows fragments obtained from a plasmid digested with the restriction enzymes X and Y. The fragments cut by enzyme X alone are in Lane 1; the fragments cut by enzyme Y alone are in Lane 2; and the double digest using both enzymes X and Y is in Lane 3.

17. The plasmid diagram below shows three restriction sites represented by slashes through the circle. Each of the three restriction sites are for either Enzyme X or Enzyme Y. You must determine by looking at the gel which of the three cuts would be made by Enzyme X, and which would be made by Enzyme Y. The numbers within the plasmid indicate the size of the nucleotide fragment generated by cutting the plasmid.

Cuts 1, 2, and 3 would bemade by which enzymes, respectively?

  1. X, Y, Y b. Y, X, Y c. Y, Y, X d. X, X, Y

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