Recombinant DNA Technology and Genomics Exam Questions and Answers Already Passed
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Recombinant DNA Technology and Genomics
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Recombinant DNA Technology And Genomics
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 ...
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
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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