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Question
Explain the properties of a good or ideal cloning vector for rDNA technology.
Solution
Following characteristic properties, a cloning vector must possess in order to be used in rDNA technology:
- A good vector should have the ability of independent replication so that as the vector replicates (through ori gene) and a large number of copies of the DNA insert will be formed.
- The vector should be able to easily introduce into host cells.
- A vector should have marker genes for antibiotic resistance.
- A vector must contain a unique cleavage site in one of the marker genes for the restriction enzyme.
- It should have at least suitable control elements like a promoter, operator, ribosomal binding sites, etc.
- The plasmids obtained naturally do not possess all the characteristics. Hence, they are constructed by inserting a gene for antibiotic resistance. eg. pBR322, pBR320, pACYC177 are the constructed plasmids. pBR322 is mostly used in rDNA technology in plants.
RELATED QUESTIONS
Rearrange the following in the correct sequence to accomplish an important biological reaction:
(a) In vitro synthesis of copies of DNA of interest
(b) Chemically synthesized oligonucleotides
(c) Enzyme DNA-polymerase
(d) Complementary region of DNA
(e) Genomic DNA template
(f) Nucleotides provided
(g) Primers
(h) Thermostable DNA-polymerase (from Thermus aquaticus)
(i) Denaturation of ds-DNA
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i. DNA is negatively charged molecule and so it is loaded on gel towards the Anode terminal.
ii. DNA fragments travel along the surface of the gel whose concentration does not affect movement of DNA.
iii. Smaller the size of DNA fragment, larger is the distance it travels through it.
iv. Pure DNA can be visualized directly by exposing to UV radiation.
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Write a short note on vectors.
