[Cloning, expression and prediction of protein structure of Ts3 gene of Cysticercus cellulosae].
OBJECTIVE
To discover a new gene of candidate molecule for the diagnosis of cysticercosis.


METHODS
Cysticercus cellulosae cDNA library was immunoscreened with pooled serum of cysticercosis patients. The encoded protein of a new gene, Ts3, was analyzed through biology software (EXPASY) on line. The new gene was cloned, the prokaryotic expression vector pET28a (+)-Ts3 was constructed, and the expressed result was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting.


RESULTS
The new gene Ts3 (GenBank accession No. EU338456) was obtained with 531 bp and an ORF of C. cellulosae. With relatively stable structure and physicochemical properties, the new gene was anon-transmembrane protein and contained more protein kinase C phosphorylation site. To induce its expression, the Ts3 gene was cloned into the vector pET28a (+), and a protein with a relative molecular mass Mr21000 was obtained which was recognized by the sera of cysticercosis cases.


CONCLUSIONS
The Ts3 gene of C. cellulosae was expressed, and the protein shows adequate antigenicity.
Zheng
Sheng-sheng
SS
Department of Parasitology, Anhui Medical University, Hefei 23003, China.
Sun
Jun-min
JM
Xia
Zi-huan
ZH
Ye
Wei
W
Yang
Jing-hua
JH
Wang
Xue-long
XL
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003552:Cysticercus / Q000235:genetics* / Q000276:immunology; D015723:Gene Library; D017238:Genes, Helminth; D015801:Helminth Proteins / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology
D015801:Helminth Proteins; D011993:Recombinant Fusion Proteins