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- W2348524811 abstract "Objective To express and identify liver cancer cells-associated CK-1 gene of Trichinella spiralis in prokaryotic cells. Methods CK-1 gene was amplified by PCR using the cDNA of T. spiralis as a template and cloned into prokaryotic expression vector pET-28a(+). The constructed recombinant plasmid pET-28a-CK-1 was transformed to E. coli BL21(DE3) which was induced with IPTG for 3,4 and 5 h respectively. The expressed product was analyzed by 12% SDS-PAGE. Mouse antisera were prepared by immunizing mice with purified T. spiralis muscle larvae,H7402 cells and recombinant CK-1 protein,and used as the first antibody for Western blot respectively. Results Restriction analysis and sequencing proved that recombinant plasmid pET-28a(+)-CK-1 was constructed correctly. The optimal time for induction with IPTG was 4 h. The expressed recombinant CK-1 protein,in a form of inclusion body,contained 37. 5% of total somatic protein. Recombinant CK-1 protein was recognized by positive mouse sera against T. spiralis muscle larvae and against whole H7402 cells,with specific bands with relative molecular mass of 38 500,which was proved as a cross antigen with good reactogenicity. Conclusion The CK-1 of T. spiralis was successfully expressed in prokaryotic cells,which laid a foundation of further study on the anti-tumor effect of T. spiralis." @default.
- W2348524811 created "2016-06-24" @default.
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- W2348524811 date "2014-01-01" @default.
- W2348524811 modified "2023-09-28" @default.
- W2348524811 title "Prokaryotic expression and identification of liver cancer cells-associated CK-1 gene of Trichinella spiralis" @default.
- W2348524811 hasPublicationYear "2014" @default.
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