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1. The most recent techniques developed in the biological sciences allow the manipulation of DNA with the ultimate goal of intervening directly with the _______ fate of organisms. genetic
2. Small circular, extrachromosomal DNA segments are known as ________. plasmids
3. Enzymes that cleave DNA at specific sites are called ______________. restriction endonucleases
4. The procedure for producing a line of genetically identical cells from a single "altered" cell is called _______. cloning
5. Single-stranded complementary tails that are produced by restriction digestion are called ______ ends. sticky
6. Viruses and bacterial plasmids are used as ______ to insert foreign DNA into host cells and create recombinant genomes. vectors
7. Gel ________ is a process that separates DNA or protein fragments according to their size, by causing them to migrate within a gel in response to an electric field. electrophoresis
8. The _________ from Agrobacterium tumefaciens is used in genetic engineering involving crop plant genes. Ti plasmid
9. _________ is a human protein synthesized in small amounts that can dissolve blood clots. TPA
10. A ______ vaccine is produced by using only a part of the viral genome. subunit
11. In addition to bacteria, _______ can also be used as vectors to insert foreign DNA into host cells and create recombinant genomes. viruses
12. Small, circular auxiliary DNA molecules of bacteria which are commonly used in biotechnology are referred to as plasmids.
13. DNA can be cleaved at specific sites, generating smaller DNA fragments with short single-stranded ends. The chemical tool used to cleave the DNA is called a(n) restriction enzyme.
14. Single-stranded ends generated by the same restriction enzyme are complementary to each other. The can be joined together, even when the source of the DNA is different.
15. Two strands of DNA that have been cut by an endonuclease can be sealed together by a(n) ligase enzyme.
16. A search for sequences that are complementary to the desired sequence of a DNA fragment uses a technique called hybridization.
17. All of the following involve molecular biology techniques except using any cell as a vector.
18. Plasmids must have two components, an origin of replication to allow it to replicate in E. coli
independently of the chromosome, and a selectable marker, usually antibiotic resistance.
19. DNA fragments complementary to the DNA being investigated are referred to as
20. Restriction enzymes are endonucleases.
21. An enzyme that has been isolated from retroviruses is reverse transcriptase.
22. Restriction enzymes recognize DNA sequences that are palindromes.
23. The ends of DNA fragments produced by the same restriction enzyme can be joined together by another enzyme, DNA ligase.
24. Vectors are vehicles of transport for foreign DNA fragments into host cells, and are usually either a virus or a bacterial plasmid.
25. Most genetic engineering experiments include four stages. Which of the following is not one of them? integration of the entire bacterial chromosome
26. The most common way to follow bacterial transformation with a plasmid is by conferring antibiotic resistance.
27. The polymerase chain reaction (PCR) includes all of the following steps except addition of new Taq polymerase after each cycle.
28. In genetic engineering experiments, a common way of identifying a transformed cell is by using a "probe," which is a specific complimentary single stranded DNA.
29. When electrical current is applied during a gel electrophoresis procedure, the DNA fragments are separated by the size of the fragments.
30. If the lacZ gene is functional it produces an enzyme known as _____________,which allows the bacterial cell to metabolize the sugar, X-gal. beta-galactosidase
31. A commercially significant human protein now produced in bacteria is human insulin.
32. A polymerase chain reaction (PCR) involves three steps. The correct order of those steps is denaturation, annealing of primers, and then primer extension.
33. A plasmid used to carry genes into crop plants is Ti.
34. Crop plants have been protected from certain insects by the addition of a gene for a specific insect toxin from the bacterium Bacillus thuringiensis.
35. Scientists can distinguish between DNA of different individuals, thus making this information useful in criminal investigations. The technique used is called restriction fragment length polymorphisms.
36. The development of a subunit vaccine for herpes and hepatitis B has potential in the future. The proposed subunit vaccine would incorporate part of the protein—polysaccharide coat of either the herpes or hepatitis B with the cowpox virus.
37. Some of the useful applications of genetic engineering include all of the following except to clone the perfect human being.
38. Agrobacterium is more difficult to use in genetic engineering procedures conducted on cereals such as corn, rice, and wheat because it does not infect these types of plants.
39. Plant scientists have successfully engineered some broadleaf crop plants to be resistant to the application of glyphosate. Glyphosate works by inhibiting an enzyme known as EPSP synthetase, which is required for aromatic amino acid production.
40. An insecticidal protein has been discovered in a bacterium known as Bacillus thuringinensis.
41. Golden rice was genetically engineered by Ingo Potrykus in Switerland by combining genes from other organisms into the genome of common white rice. Which organism from the list below was NOT used? Agrobacterium
42. DNA restriction enzymes are labeled I and II. Type I restriction enzymes make simple cuts across both DNA strands.
43. A vector "carries" ______________ into a host cell. a recombinant DNA molecule
44. Autonomous auxiliary DNA circles found in bacteria; easily enter the organism they are found in; used as a vehicle to transport desired foreign genes. plasmids
45. Technique used to amplify a desired piece of DNA. PCR
46. Technique used to analyze DNA products after restriction enzyme digestion; "fingerprints." RFLP
47. Organisms produced by moving genes of one species to another; desired characteristics such as disease resistance or increased size are transferred. transgenic organisms
48. Ends of DNA fragments produced by asymmetric cleavage with restriction endonucleases; same ends are produced by the same enzyme irrespective of the source of the DNA. sticky ends
49. Bam HI is a Type II restriction enzyme that recognizes the sequence 5' GGATCC 3', and cuts between the two Gs, leaving a 5' overhang. If you were to digest DNA with Bam HI, what would be the sequence of the overhanging sticky end of the complimentary 5' GATC 3'
50.A 6.85 kb Eco RI fragment of DNA is shown below. The locations of several restriction sites are indicated. Scale is approximate.If you were to digest this fragment of DNA with Pst I, how many pieces of DNA would you obtain? 3
51. If you were to perform a double digest of this DNA using Bam HI and Eco RI, what is the size of the DNA fragment that would migrate the fastest on an agarose gel? 1038 kb
52. You are performing a Southern blot analysis. To obtain a probe, you digest the above piece of DNA with Pst I and isolate a 2.6 kb Pst I fragment. You radiolabel the fragment and use it to probe a Southern blot of the above DNA that has been digested w 1038 kb, 3529 kb
53. Which of the following vectors is the best choice to clone a 23 kb genomic DNA fragment? phage vector
54. You are trying to clone a piece of DNA into a plasmid vector that exhibits blue-white selection. Following ligation and transformation, you plate the host cells onto several plates containing the appropriate selective media. The next day you examine t 43 white colonies, 6 blue colonies
55. Your research project involves the characterization of a signal transduction pathway, and you would like to identify proteins that may interact with one of the pathway components. Which of the following libraries could you screen for this purpose? an expression library
56. When compared with non-cancerous cells of the same tissue, cancer cells exhibit a number of changes in gene transcription. To identify genes that are differentially expressed in cancer cells versus noncancer cells, one could perform subtractive hybridization.
57. A researcher identified a new enzyme and is interested in determining the mRNA expression pattern of this gene. Which of the following techniques would accomplish this goal? northern blot
58. You are hired as a scientific consultant for a popular television show about forensic analysis of crime scene evidence. You are told that in an upcoming scene, investigators will compare the DNA profiles of a known suspect and an unknown biological sa DNA fingerprinting
59. Which of the following types of information would be most useful in an effort to move a DNA fragment from one plasmid vector to another, using molecular cloning techniques? restriction maps of the plasmids
60. Which of the following two enzymes are needed to construct a cDNA library reverse transcriptase and DNA ligase
61. In a manual enzymatic DNA sequencing reaction, the nucleotide ddT is incorporated randomly at a single T position in the synthesized DNA strand, leading to termination of the reaction.
62. Automated enzymatic sequencing employs different colored fluorescent dyes conjugated to dideoxy terminators and capillary gel
63. The development of PCR has revolutionized molecular biology and has led to numerous advances in basic research, as well as in medicine and forensic science. Which of the following areas does not relyon PCR? generation of hybrid plants
64. You are performing a yeast two hybrid screen to identify protein-protein interactions. Your bait is a 25 kD protein fused to the GAL4 DNA-binding domain. Which of the following best describes the nature of the library that you are using? The library contains the prey fused to the GAL4 activation domain.
65. To generate a transgenic mouse, one would introduce the knocked-out gene into embryonic stem cells, inject the embryonic stem cells into a blastocyst stage embryo, implant the injected blastocyst into a pseudopregnant female.
66. Which of the following is not an example of reverse genetics? identification of the gene responsible for particular mutant phenotype
67. A vaccine that depends on the cellular immune response of killer T cells is called a DNA vaccine.
68. Current vectors used for gene therapy insert into the genome as a random event. All of the following are potential problems that may result from this method of integration except expression of the transgenic gene.
69. What of the following is a potential disadvantage of using transgenic plants for vaccine production? risk of contamination with herbicides and pesticides
70. What is the environmental benefit of the Enviropig? reduction of phosphate excretion in pig waste
71. Some food crops are difficult to grow for a variety of reasons. For example, strawberries are easily susceptible to frost damage. How could one use genetic engineering techniques help to overcome this type of problem, and which type of animal or plant To increase the frost hardiness of strawberries, one could generate a strain of transgenic strawberries using a gene that confers frost resistance. To identify such a gene, one could look to an animal or plant that lives in a very cold environment. Such o
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