Escherichia coli pTUC202 plasmid原核表达载体质粒 BioVector NTCC质粒载体菌种细胞基因保藏中心

Escherichia coli pTUC202 plasmid原核表达载体质粒

pTUC202 plasmid harbors the native gene cluster for the production of MccJ25 while pJP3 contains an engineered mcjABCD gene cluster in which the mcjA gene is placed under the control of an IPTG-inducible promoter (Fig. 2). The pJP4 plasmid is identical to pJP3 except that it lacks the mcjA gene. Both pJP3 and pJP4 are derivatives of the commercial vector pQE-60 (Qiagen). To construct each plasmid encoding a mutant mcjA gene, the mcjA signal sequence was first amplified from pTUC202 using primers mcjA signal F and mcjA signal R. The mcjA mature sequence with the designed mutations was synthesized by primer overlap PCR using the forward and reverse primers for each mutant listed in Table I. The amplified signal sequence was then overlapped with the mature sequence in a second PCR step to generate a reconstituted mutant mcjA gene.

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