my vector contain ccdB gene in MCS, can i use this as mock control during transf - (Feb/09/2007 )
i subclone my gene for expression into invitrogen pcDNA-Dest40 vector, which contain ccdB gene in MCS to facilitate positive selection after transformation of E.coli. this ccdB gene is lethal to bacteria, so only those with insert disrupted vector will grow.
but when i transfect the vector to hepatocelluar cancer cell, can i use the original pDEST40 as mock control? i do not know if this ccdB gene is cytotoxic to hepatocelluar cancer cell. my boss tell me this will not affect a lot. but if my target gene is not so powerful, i can not distinguish between test and mock.
or can i use a irrelveant vector? such as pcDNA3.1 as control? the vector self should not have any effect if they did not contain expression reading frame, is this correct?
We use pcDNA3.1 for mock control.