[Cloning, expression and identification of membrane protein Tetraspanin 2-A of Schistosoma japonicum].
OBJECTIVE
To clone and express a membrane protein (Tetraspanin 2) gene of Schistosoma japonicum (SjTsp2).


METHODS
A pair of primers was designed to amplify the SjTsp2 gene which was subcloned into prokaryotic plasmid pET28a(+). The recombinant plasmid was transformed into E. coli BL21(D3) and followed by expression of the protein induced by IPTG. The protein was purified by affinity chromatography and used to immunize BALB/c mice. Dilution of antibody against SjTsp2 was determined by ELISA. The protein was also identified by Western blotting.


RESULTS
Big loop of SjTsp2-A, 228 bp, was amplified in vitro by PCR. Its deduced amino sequence shared 52% similarity with SmTsp2. The soluble recombinant SjTsp2-A was expressed in the experiment and high dilution antibody against the recombinant (1:32,000 in maximum) was produced in immunized mice. SjTsp2-A reacted positively with sera of acute and chronic schistosomiasis patients but not with sera from healthy persons by Western blotting.


CONCLUSIONS
SjTsp2 has been expressed and shows certain antigenicity.
Wang
Yang-Yang
YY
Department of Microbiology and Parasitology, Anhui Medical University, Hefei 230032, China.
Liu
Miao
M
Zhu
Shao-Chun
SC
Ren
Cui-Ping
CP
Shen
Ji-Jia
JJ
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D000947:Antigens, Helminth / Q000235:genetics / Q000276:immunology; D015153:Blotting, Western; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D015723:Gene Library; D015801:Helminth Proteins / Q000235:genetics* / Q000276:immunology; D007114:Immunization; D008565:Membrane Proteins / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000276:immunology; D012549:Schistosoma japonicum / Q000235:genetics* / Q000276:immunology; D012554:Schistosomiasis japonica / Q000097:blood / Q000469:parasitology
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D015801:Helminth Proteins; D008565:Membrane Proteins; D011994:Recombinant Proteins