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Recombinant DNA Technology and Genomics Exam Questions and Answers Already Passed

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Recombinant DNA Technology and Genomics Exam Questions and Answers Already Passed Clone - Answers a molecule, cell, or organism that was produced from another single entity Restriction Enzymes - Answers DNA cutting enzymes (molecular scissors) -Primarily found in bacteria -Cut DNA by cleaving ...

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  • October 21, 2024
  • 7
  • 2024/2025
  • Exam (elaborations)
  • Questions & answers
  • Recombinant DNA Technology and Genomics
  • Recombinant DNA Technology and Genomics
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Recombinant DNA Technology and Genomics Exam Questions and Answers Already Passed

Clone - Answers a molecule, cell, or organism that was produced from another single entity

Restriction Enzymes - Answers DNA cutting enzymes (molecular scissors)

-Primarily found in bacteria

-Cut DNA by cleaving the phosphodiester bond that joins adjacent nucleotides in a DNA strand

-There are 4 or 6 bp cutters because they recognize restriction sites with a sequence of 4 or 6
nucleotides

Plasmid DNA Vectors - Answers circular form of self-replicating DNA

•Can be manipulated to carry and clone other pieces of DNA

Plasmid DNA - small circular pieces of DNA found primarily in bacteria

•Are considered "extra-chromosomal" DNA because they are in the cytoplasm in addition to the
bacteria chromosome

•Are small: approx. 1 - 4 kb

•Can replicate independently of chromosome

restriction site - Answers Restriction Enzymes Bind to, recognize, and cut DNA within specific sequences
of bases called a

Each restriction site is a palindrome - reads same forward and backward on opposite strands of DNA

Advantage of enzymes that produce sticky ends - Answers Preferred for cloning because DNA fragments
with sticky ends can be easily joined together because they base pair with each other by forming weak
hydrogen bonds

vectors - Answers pieces of DNA that can accept, carry, and replicate other pieces of DNA

Recombinant DNA Advisory Committee - Answers 1975 NIH formed the RAC

Purpose of RAC: evaluate recombinant technology and establish guidelines for research

-1976 RAC published set of guidelines for working with recombinant organisms

Transformation of Bacterial Cells - Answers -Very inefficient process

-A process for inserting foreign DNA into bacteria

•Treat bacterial cells with calcium chloride

, •Add plasmid DNA to cells chilled on ice

•Heat the cell and DNA mixture

•Plasmid DNA enters bacterial cells and is replicated and express their genes

Electroporation - Answers Apply brief pulse of high voltage electricity to create tiny holes in the bacteria
cell wall that allow the DNA to enter

Selection - Answers is a process designed to facilitate the identification of recombinant bacteria while
preventing the growth of non-transformed bacteria and bacteria that contain plasmid without foreign
DNA

Antibiotic selection - Answers plate transformed cells on plates containing different antibiotics to
identify recombinant bacteria and non-transformed bacteria

-Does not select for plasmid containing foreign DNA vs. recircularized plasmid

Blue-white selection - Answers -DNA is cloned into the restriction site in the lacZ gene

-When it is interrupted by an inserted gene, the lacZ gene cannot produce functional beta gal

-When Xgal (artificial lactose) is added to the plate, if functional lacZ is present = blue colony

-Non-functional lacZ = white colony = clone = genetically identical bacterial cells each containing copies
of recomb. plasmid

insulin - Answers First human protein expressed via recombinant techniques

•Clone human insulin DNA sequence into a plasmid and the bacteria cells were then used to synthesize
the protein product of the cloned gene

•Can generate lots of pure protein via this technique

Bacterial Plasmid Vectors (circular) - Answers 6-12kb

Applications: DNA cloning, protein expression, sub-cloning, direct sequencing of insert DNA

Disadvantages: Restricted insert size, limited expression of proteins, copy number problems; replication
restricted to bacteria

Bacteriophage (linear) - Answers 25kb

Applications: cDNA, genomic and expression libraries

Disadvantages: packaging limits DNA insert size; host replication problems.

Cosmid (circular) - Answers 35kb

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