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Is it possible to make your own Rosetta(DE3) pLysS by transforming pLysS-Rosetta


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#1 benz42428

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Posted 28 July 2012 - 02:47 PM

I am planning to express my protein in Rosetta strain. Instead of purchasing the commercially available Rosetta(DE3) pLysS strain, my PI told me to transform BL21(DE3) with the pLysS-Rosetta plasmid that he mini prepped from the Rosetta strain he borrowed from another lab. I wonder if this is going to work as well as using the commercially available Rosetta strain, given that one of my friends told me the strategy that my pI urged me to use didn't work very well for him. Any thoughts?

Also, if this strategy happens to work, can I make a Rosetta strain out of C43(DE3) cells? Thanks

#2 pDNA

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Posted 29 July 2012 - 12:27 AM

why not? ...if the plasmid has been prepared from a valid source and if its identity has been confirmed ...then it should be a possible way to go!

The only thing is that there are different Rosetta strains circulating ...Rosetta and Rosetta 2 ...then i would ask again what strain has been used to isolate the plasmid. If no one really knows i would assume that the source is not valid and order a new batch of cells! Because it is a pitty when people work on "unconfirmed" plasmids ...if you don't know what it is exactly ...then screw it!

Regards,
p




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