Cloning And Expression Of Heat-Labile Enterotoxin B Subunit And The Toxicity Experimentation In Vivo

PARTâ… Cloning and Expression of pBV220-LTBObjective: To construct and express pBV220-LTB,we distill Heat-labile enterotoxin B subunit from E.coli H 44815 by gene-engineering, which would lay a foundation for the biological idiosyncrasy.Methods: Heat-labile enterotoxin B subunit gene was amplified by PCR and cloned into plasmid pBV220. The recombinant plamsid was identified by sequencing ,then transformed into E.coli DH5a.. The transformant colony was induced with IPTG. Purify the expressed protein by Ni²⁺-NTA column chromatography. Expression of the protein was analyzed by SDS-PAGE .Results: The gene fragment at length of 396 bp was amplified and successfully cloned into plasmid pBV220.SDS-PAGE showed a protein band with relative molecular weight of 30 000,which was consistent with the expectation.Conclusion: The recombinant plasmid of pBV220-LTB was successfully constructed and Heat-labile enterotoxin B subunit gene was highly expressed in E.coli DH5a.PARTâ…¡The rabbit`s toxicity experimentation of the protein of Heat-labile enterotoxin B subunit...