I cloned my gene of interest into pCMV tag2c, pGEX-3X, pLXSN, VenusC1 and VenusN1. From DNA sequencing, all clones were correct and inframe. I can't express my protein from pCMV tag2C and pLXSN; however, VenusC1, N1 and pGEX-3x expressed very well in RHF cells and BL21 respectively. Could anyone please give me suggestions ???
Western blot was used to detect expression
293T cells were used for transfection
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No expression in 293T cells
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