[Study on expression of curcin gene cloned from Jatropha curcas in Escherichia coli].
OBJECTIVE
To construct mature protein curcin gene prokaryotic expression vectors in Escherichia coli and choose the optimal inducing condition of the recombinant strains.


METHODS
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.


RESULTS
The gene encoding of mature protein curcin was amplified by PCR and the recombinant strains PRM and PRB were obtained.


CONCLUSIONS
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.
Luo
Mengjun
M
Family Planning Technology Institute of Chengdu, Chengdu 610031, China.
Chen
Fang
F
Yan
Fang
F
Liu
Weixin
W
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression / Q000187:drug effects; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D018745:Genome, Plant / Q000235:genetics; D031286:Jatropha / Q000235:genetics*; D054789:Ribosome Inactivating Proteins, Type 1 / Q000096:biosynthesis / Q000235:genetics*; D013696:Temperature
D054789:Ribosome Inactivating Proteins, Type 1; C515388:curcin protein, Jatropha curcas