Retroviral vector and recombination - Are Retroviral vector plasmids susceptible to recombination?why? (Apr/16/2007 )
Recently I heard that retroviral vector plasmids like pMSCV can be easily recombined with bacterial genomic DNA so I can not use bacteria strains like DH5a for transformation. Does anyone know why;
1. retroviral vector plasmids can be easily recombined?
2. it is not good idea to use bacterial strains like DH5a to amplify retroviral plasmid?
Thank you so much in advance!
don't know your specific plasmid, but mosly retroviral vectors have LTR-sequences, that are repetitive. E. coli standard doesn't have that sort of repetitive sequences, so they can get "kicked out" by recombination.
There are certain strains (stbl from invitrogen and SURE from stratagene, probably other companies have some strains too) that are recombination defective, so the recombination won't happen.
I grow my retroviral vectors @ 30°C with shaking at 180 rpm and have had no problems with DH5a (growing takes longer though).