[Immunoscreening of schistosomulum cDNA library of Schistosoma japonicum and preliminary identification of membrane-associated protein abundant in tegument].
OBJECTIVE
To obtain genes encoding the novel molecules for diagnosis of schistosomiasis.


METHODS
Juvenile S. japonicum cDNA library was immunoscreened to obtain positive clones. By DNA sequencing and sequence analysis, the target gene was amplified by PCR and subcloned into prokaryotic plasmid pET28a. The recombinant plasmid was transformed into E. coli BL21 followed by expression of the protein induced by IPTG. The protein was identified by Western blotting.


RESULTS
34 positive clones were obtained, 24 of which were chosen to be sequenced, 13 of which were Sj22600 gene. The protein were recognized with sera of infected rabbits and patients with acute and chronic schistosomiasis by Western blotting.


CONCLUSIONS
The gene coding for Sj22600 membrane protein was screened with high frequency in the cDNA library. The E. coli BL21 transformed with the recombinant plasmid can express the fusion protein, which shows immunoactivity.
Zhu
Shao-chun
SC
Department of Microbiology and Parasitology, Anhui Medical University, Hefei 230032, China.
Zhao
Zhi-rong
ZR
Lei
Li
L
Zhang
Lin-jie
LJ
Shen
Ji-jia
JJ
D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western / Q000379:methods; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D015723:Gene Library; D015801:Helminth Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011817:Rabbits; D012549:Schistosoma japonicum / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D017422:Sequence Analysis, DNA; D044967:Serum / Q000276:immunology
D018076:DNA, Complementary; D015801:Helminth Proteins; D008565:Membrane Proteins