[Expression of proteinase cathepsin L1 gene of Schistosoma japonicum in Escherichia coli].
Bing Yi, Zhuoya Li, He Ai, Xiaoying Zheng, Bin Zheng, Haojie Zhou, Yi Wang, Ruilin Zhang, Nan Yu, Ximei Zhan
Abstract
Bing Yi, Zhuoya Li, He Ai, Xiaoying Zheng, Bin Zheng, Haojie Zhou, Yi Wang, Ruilin Zhang, Nan Yu, Ximei Zhan
Abstract
OBJECTIVE: To express the proteinase cathepsin L1 gene of Schistosoma japonicum (SjCL1) in Escherichia coli JM109 cells. METHODS: The SjCL1 gene was amplified from the recombinant plasmid pcDNA3-SjCL1 by PCR. The gene was cloned into a prokaryotic expression vector pGEX4T-1 to construct a recombinant plasmid pGEX-SjCL1. The E. coli JM109 cells were transformed with the recombinant plasmid pGEX-SjCL1 and the transformants were induced by IPTG to express the recombinant protein, the target protein was then identified by SDS-PAGE and Western blotting. RESULTS: A 1 kb length PCR product was obtained and a recombinant plasmid pGEX-SjCL1 was constructed. The expression product was detected by SDS-PAGE and Western blotting and an expression band about 62000 was found. CONCLUSION: The SjCL1 gene is effectively expressed in the E. coli JM109 cells.
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OBJECTIVE: To express the proteinase cathepsin L1 gene of Schistosoma japonicum (SjCL1) in Escherichia coli JM109 cells. METHODS: The SjCL1 gene was amplified from the recombinant plasmid pcDNA3-SjCL1 by PCR. The gene was cloned into a prokaryotic expression vector pGEX4T-1 to construct a recombinant plasmid pGEX-SjCL1. The E. coli JM109 cells were transformed with the recombinant plasmid pGEX-SjCL1 and the transformants were induced by IPTG to express the recombinant protein, the target protein was then identified by SDS-PAGE and Western blotting. RESULTS: A 1 kb length PCR product was obtained and a recombinant plasmid pGEX-SjCL1 was constructed. The expression product was detected by SDS-PAGE and Western blotting and an expression band about 62000 was found. CONCLUSION: The SjCL1 gene is effectively expressed in the E. coli JM109 cells.
Key concepts: Recombinant DNA, Plasmid, Molecular biology, Schistosoma japonicum, Escherichia coli, Biology, lac operon, Gene