pMV261-sfgfp plasmid vector分支桿菌綠色熒光表達(dá)載體質(zhì)粒

Gene encoding superfolder GFP (sfGFP) was amplified from pKD4-sfGFP plasmid with specific primers which were cloned in pMV261 and pVV16 vectors at either EcoRI/HindIII or only HindIII restriction sites. Thus, creating pMV261::sfGFP and pVV16::sfGFP. Rv0646c and MSMEG_1352 ORFs were amplified by PCR with specific primers  containing NdeI and BamHI restriction sites in 5′ and 3′ ends, respectively. Digested and purified products were cloned into the pVV16::sfGFP in frame with the sfGFP gene and thus generating a translational fusion. M. smegmatis was electroporated with 1.5 μg of pMV261::sfGFP and these latter constructs; recombinant clones were selected on to specific medium and overexpression of the two chimeric proteins were checked by immunoblotting as described above.

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