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China Journal of Chinese Materia Medica ; (24): 656-659, 2009.
Article in Chinese | WPRIM | ID: wpr-265336

ABSTRACT

<p><b>OBJECTIVE</b>To construct mature protein curcin gene prokaryotic expression vectors in Escherichia coli and choose the optimal inducing condition of the recombinant strains.</p><p><b>METHOD</b>The gene encoding of curcin was amplified from the genome of Jatropha curcas seeds by PCR and cloned into the expression vectors pQE-30 and pET-32 obtaining recombinant vectors pQE-R and pET-R respectively. The two vectors were transferred into E. coli BL21 (DE3) and the recombinant strains PRM and PRB were attained respectively. PRM and PRB were induced by different revulsants and under different temperature and time.</p><p><b>RESULT</b>The gene encoding of mature protein curcin was amplified by PCR and the recombinant strains PRM and PRB were obtained.</p><p><b>CONCLUSION</b>The results showed that PRB could not produce recombinant protein under such conditions. However, PRM could highly produce recombinant protein induced by 0.5 mmol x L(-1) IPTG at 28 degrees C for 6 h.</p>


Subject(s)
Cloning, Molecular , Escherichia coli , Genetics , Gene Expression , Genetic Vectors , Genetics , Metabolism , Genome, Plant , Genetics , Jatropha , Genetics , Ribosome Inactivating Proteins, Type 1 , Genetics , Temperature
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