To obtain the recombinant core domain of porcine zone pellucida 3β （cZP3β） for the further research on its functions Methods The nucleotide sequence region from 44 to 306 codons of pZP3β entire eDNA was obtained ...To obtain the recombinant core domain of porcine zone pellucida 3β （cZP3β） for the further research on its functions Methods The nucleotide sequence region from 44 to 306 codons of pZP3β entire eDNA was obtained by PCR and then was cloned into pET-3c vector. After being identified, recon was transformed into E.coli BL21 （DE3） pLysS and then induced by IPTG. Results The recombinant cZP3β was expressed in E. coli up to 15% of total cellular proteins, and was made sure by Western blot analysis. Conclusion The research on expression of core domain of pZP3β could benefit to further investigation of its immunogenicity and the development of antigen preparation.展开更多
Objeetive To obtain the recombinant non fusion extracellular porcine zona pellucida protein 3β （pZP3β） in E.coli Methods By modificated the transition initiation region （TIR） in primers, synthetic nucleotide was...Objeetive To obtain the recombinant non fusion extracellular porcine zona pellucida protein 3β （pZP3β） in E.coli Methods By modificated the transition initiation region （TIR） in primers, synthetic nucleotide was gained by PCR. Such gene was cloned into pET-3c vector and transformed into E.coli BL21（DE3）pLysS. Results The recombi.ant nonfusion extracelhtlar pZP3β was expressed in E.coli to 10% of total cellular proteins, and identified by the Western blot method. Conclusion Modification of nucleotide without changing amino acid sequences is an effective means to increase non fusion expression rate of recombinant proteins, such as pZP3β in E. coll.展开更多
基金This study was supported by the Science & Technology Plan (No. 2001C12001) of Guangdong Province,P.R. China
文摘To obtain the recombinant core domain of porcine zone pellucida 3β （cZP3β） for the further research on its functions Methods The nucleotide sequence region from 44 to 306 codons of pZP3β entire eDNA was obtained by PCR and then was cloned into pET-3c vector. After being identified, recon was transformed into E.coli BL21 （DE3） pLysS and then induced by IPTG. Results The recombinant cZP3β was expressed in E. coli up to 15% of total cellular proteins, and was made sure by Western blot analysis. Conclusion The research on expression of core domain of pZP3β could benefit to further investigation of its immunogenicity and the development of antigen preparation.
基金This study was supported by the Science & Technology Plan (No. 2001C12001) of Guangdong Province,P.R. China
文摘Objeetive To obtain the recombinant non fusion extracellular porcine zona pellucida protein 3β （pZP3β） in E.coli Methods By modificated the transition initiation region （TIR） in primers, synthetic nucleotide was gained by PCR. Such gene was cloned into pET-3c vector and transformed into E.coli BL21（DE3）pLysS. Results The recombi.ant nonfusion extracelhtlar pZP3β was expressed in E.coli to 10% of total cellular proteins, and identified by the Western blot method. Conclusion Modification of nucleotide without changing amino acid sequences is an effective means to increase non fusion expression rate of recombinant proteins, such as pZP3β in E. coll.
