c. True or False (circle T or F. [For Extra credit, correct they are falsel. fal

Question

c. True or False (circle T or F. [For Extra credit, correct they are falsel. false statements or explain why 32. Unlike DNA replication in the cell, PcR uses heat to separate DNA strands. (T/F 33. Restriction endonucleases are enzymes that cut ONA at specific double-stranded sequences min Taq polymerase, an enzyme found in thermophilic bacteria, is used to replicate DNA in PoR (T/Fl 34, ages, viruses that infect bacteria, can be used to deliver recombinant DNA into host cell. /F 35. 36. Antibiotic resistonce genes are often included in cloning vectors to serve as selection merkers, IT/F 37 Restriction Fragment Length Polymorphisms (RFLP) can be used to generate DNA fingerprints (T/Fl 38. Green Fluorescent Protein (GFP) is an example of a "reporter protein" naturally found in bacteria. (T/Fl 39 Reverse transcriptase is an enzyme that generates "sticky ends." (T/Fl segment of a plasmid vector that contains multiple restrietion enzyme sites. (T/F 40. A polylinker is a 41. Antibiotic resistance genes are used to identify bacterial colonies containing recombinant DNA T/F) 43. The gene that encodes GFP is frequently used as reporter gene. (T/F Agarose gel electrophoresis is a technique used to separate DNA fragments by size. 44, 45. Transgenic animals are produced using recombinant DNA technology. (T/F p, Multiple Choice 46. Which of the following IS NOT required for PCR? a. DNA Polymerase d. Primers e. None of the above (all are required) 47, Recombinant DNA: a. ls defined as a mix of human and animal DNA. b. Contains a segment of DNA from a foreign source. c. created from embryonic stem cells. d. required for human cloning. e. None of the above.

Explanation / Answer

Answer:

(32) True

DNA replication in cells uses helicase. In PCR this step is performed by heating the sample to 95C.

(33) True

Restriction endonucleases are enzymes whivh cut ds-DNA at or near specific site. Type 2 restriction endonucleases cut at the specific site and widely osed in recombinant DNA technology. example: EcoR1.

(34) True

Taq polymerase is isolated from Thermus aquaticus and can tolerate high temperatures. It has a optimum activity at 72C.

(35) True

Lambda phage vectors are widely used viral vector for recombinant gene delivery into host cells.

(36) True

Antibiotic resistant markers or selection markers are used to identified positive clones in culture, that is only those bacterial clones grow in antibiotic plate which has the vector with the resistant gene.

(37) True

In RFLP, restriction enzymes cleave unique sequences, that is mostly repetitive sequences in DNA which is unique for an individual resulting in a fingerprint.

(38) False

Correct statement: Green Fluorescent Protein (GFP) is an example of a reporter protein naturally found in Aequorea victoria (Jelly fish)

(39) False

Reverse transcriptase generates complementary DNA (cDNA) from mRNA. Restriction enzymes type II cleaves double stranded DNA and produces sticky ends.

(40) True

Polylinker or Multiple Cloning Site (MCS) are sequences in vectors which contain restriction sites for several restriction enzymes. These are restriction sites are unique as there is only one site per restriction enzyme in a vector.

(41) True

Antibiotic resistant genes in vectors are used as selectable marker for selecting positive bacterial clones which contain the vector with the resistant marker. The plasmids should be isolated and digested with appropriate restriction enzymes to detect for the presence of the recombinant gene.

(42) True

Human genes can be expressed in bacteria, but the encoded protein do not undergo post-translational modification since bacteria lacks this machinery. Again, the encoded protein may not fold properly and may end up in inclusion bodies.

In mice, human genes can be expressed with a common mammalian promoter (say CMV). If human specofoc promoter is used, it should be made sure that the mice model contains the appropriate transcription factors for expression.

(43) True

GFP is used as a reporter gene. In cells when the tagged protein gets expressed, GFP also gets expressed and emits green fluorescence which can be seen under confocal microscope to confirm the expression of the genes of interest.

(44) True

Agarose gel electrophoresis separates DNA by size. The smaller the size, the more is the migration of the DNA fragments in the gel.

(45) True

Transgenic animals are produced by delivering a transgene into genome of the animal through microinjection.

(46) c

Helicase is not required in PCR as this step of unwinding double stranded DNA to single strands is done by the denaturation (heating step).

(47) b

Recombinant DNA is produced by insertion of a foreign DNA in expression vectors.

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