Genomics #7 Cloning Vectors.txt

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HUSOP2014
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133283
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Genomics #7 Cloning Vectors.txt
Updated:
2012-02-10 11:37:57
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HUSOP Gen EXAM1 Cloning Vectors
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genomics cloning vectors
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  1. The main purpose of cloning DNA is?
    To make multiple copies of a unique gene sequence
  2. T/F A biological approach to cloning DNA is much less effective than chemical means.
    FALSE
  3. All of traditional DNA cloning approaches use __________ DNA structures.
    self-replicating
  4. What are the two main classes of vectors?
    • Cloning
    • Expression
  5. Name two functions of cloning vectors.
    • Modify DNA sequences within a unique segment of DNA
    • make multiple DNA or RNA copies of a specific DNA sequence
  6. What are expression vectors used for?
    To synthesize protein coded by a cloned DNA sequence.
  7. Name several types of cloning vectors and comment on the amount of DNA they can hold.
    • Plasmid (up to 5.0 kB insert),
    • bacteriophage (up to 25 kB insert),
    • phagemid (up to 25 kB insert),
    • cosmid (up to 45 kB insert),
    • YAC (yeast artificial chromosome)(100-200 kB insert)
  8. 1 kB is how many bases? What is a kBp?
    • 1000 bases,
    • 1000 base pairs
  9. The most frequently used cloning vector is the __________.
    Plasmid
  10. T/F Plasmids have double stranded, closed circular DNA with selectable markers.
    TRUE
  11. The genes encoded by bacterial plasmids include those that __________.
    Impart drug resistance
  12. pBR322 is a type of __________ vector.
    Plasmid
  13. In regards to a pBR322, what do the outer and inner cirlces represent?
    • outer represents where restriction nucleases cut the DNA
    • inner circle denotes where the resistance genes are located.
  14. T/F A different restriction nuclease is needed to open a plasmid that is used on the DNA to be cloned.
    FALSE
  15. What enzyme is used to seal cut plasmids and to add covalent bonds?
    DNA ligase
  16. T/F Bacteriophages are double stranded DNA vectors.
    FALSE
  17. Bacteriophages infect which particular type of bacteria?
    E. coli
  18. Bacteriophages contain the selectable marker __________ .
    Beta-galactosidase
  19. Transfected E. coli will stain blue when incubated in the presence of what galactosidase?
    x-gal, 5-bromo-4-chloro-3-indoyl-?-D-galactosidase
  20. A phagemid can be propogated by either a __________ or a __________.
    • Plasmid
    • bacteriophage
  21. Propagating a phagemid as a bacteriophage requires what?
    Co-infection with a bacteriophage and a phagemid
  22. Cosmids are manipulated as __________, but are expressed as __________ in the cell.
    • Bacteriophage,
    • Plasmid
  23. YAC's are propagated in which type of yeast?
    Saccharomyces cerevisiae
  24. How is cloning possible with YAC's?
    It is recognized as part of its normal complement of chromosomes.
  25. YAC's contain all of the following except:
    A) selectable marker
    B) centromere
    C) telomeric DNA
    D) Beta-galactosidase
    B-Galactosidase
  26. __________ is an endonuclease commonly found in YAC cloning vectors. What does this enzyme do?
    EcoR1- It creates sticky ends with 5' overhangs
  27. What is the difference between complementary and genomic DNA?
    • cDNA only contains only the protein coding region of a gene.
    • Genomic DNA contains the entire gene sequence.
  28. Besides the protein coding region, genomic DNA has what 4 other parts?
    • Promoters,
    • introns,
    • exons
    • 3' non-translated domain
  29. The key to cloning in eukaryotic cells is __________.
    Glycosylation
  30. __________ can accept and express proteins in DNA within different constructs.
    Cos cells
  31. What vector is used to express glycosylated proteins in insects?
    Baculovirus
  32. T/F Baculovirus can enter mammalian cells and replicate.
    FALSE
  33. Which type of vector is known as a shuttle vector?
    Bacmid

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