Cloning of Schistosoma japonicum Chinese Strain TPI Gene and Characterization of Its Expression Product in Escherichia coli.
Triose-phosphate isomerase is an important candidate for schistosoma antigens. An 800 bp DNA fragment was amplified by RT-PCR from adult Schistosoma japonicum mRNA. Sequence analysis revealed that this fragment contained S. japonicum (Chinese strain) triose-phosphate isomerase gene. Then this gene was cloned into the expression vector pGEX-4T and subsequently expressed in E. coli. The recombinant GST-fusion protein was purified by glutathione agarose affinity chromatography. Its molecular weight was determined to be 54 kD. The yield of expression was around 30 mg/L E. coli culture. Western blotting showed that the recombinant protein had good antigenicity which could be helpful for the making of anti-S. japonicum multi-valent recombinant vaccine.
Cai
Xue-Zhong
XZ
Shanghai Institute of Animal parasitology, Chinese Academy of Agricultural Sciences, Shanghai 200232, China. caassp@public.sta.net.cn
Lin
Jiao-Jiao
JJ
Yang
Guan-Zhen
GZ
Shi
Fu-Hui
FH
Shen
Wei
W
Cai
You-Min
YM
Wu
Xiang-Fu
XF