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Plasmide in recombinant DNA?

Zubnaya Feya

Combinator
10+ Year Member
5+ Year Member
Nov 13, 2008
971
3
101
Ann Arbor, MI
  1. Dental Student
guys:

what is the role of plasmide in recombinant DNA?

In order to get the gene of interest into the genome of a species researchers mostly use plasmid vectors and then viruses.

1) You make a plasmid construct (circular DNA containing gene of interest, promoter, enhancer, other elements needed for survival in bacterial cell) via restriction digestion, ligation, etc

2) You transform (transformation = taking up something inside something...) bacteria with the plasmid, let the bacteria grow - plasmid replicates with the bacterial genome.

3) You collect the replicated plasmid

4) Then you could use the plasmid in combination with packaging vectors to create a virus, transfect the cells of interest (stem cells or smth).

5) The virus gets into the cells, recombination occurs (they use lenti or retroviruses usually) and the gene of interest (that was present in the plasmid) integrates into genomic DNA of cells.

6) Then you can transfer the cells back into the organism and see the phenotype.

... I hope I didn't leave out anything.
 
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