[Prokaryotic expression and immunologic identification of P-29 gene from Echinococcus granulosus hydatid cyst].
OBJECTIVE
To clone and express P-29 gene of Echinococcus granulosus, and analyze its immunoreactivity.


METHODS
Total RNA was extracted from the hydatid cyst of E. granulosus and its P-29 gene was amplified by RT-PCR. The PCR product was cloned into pET44a(+) vector. The recombinant plasmid was transformed into E. coli BL21 (DE3) and followed by expression of the protein induced by IPTG. The protein was purified, and tested by SDS-PAGE. Its immunoreactivity was examined by Western blotting.


RESULTS
The recombinant expression plasmid was identified by PCR, double endonuclease digestion and sequencing. The recombinant Nus-P-29 was about Mr 93 000 with a concentration of 0.78 mg/ml. It was recognized by the sera of cystic echinococcosis patients.


CONCLUSIONS
The P-29 gene has been expressed with adequate immunoreactivity.
Li
Jie
J
College of Life Science, Inner Mongolia University, Key Laboratory of Mammal Reproductive Biology and Biotechnology, Ministry of Education, Hohhot 010021, China.
Wang
Zhi-gang
ZG
Hao
Xi-yan
XY
Chen
Xian-wei
XW
Wang
Hong-xia
HX
Li
Shu-yu
SY
Yang
Jiao-fu
JF
Yang
Jun
J
D000818:Animals; D000947:Antigens, Helminth / Q000235:genetics* / Q000276:immunology*; D003001:Cloning, Molecular; D048209:Echinococcus granulosus / Q000276:immunology*; D017238:Genes, Helminth; D010957:Plasmids; D011994:Recombinant Proteins / Q000276:immunology
D000947:Antigens, Helminth; D011994:Recombinant Proteins