[Cloning, expression and phylogenetic analysis of Schistosoma japonicum calcyphosine gene].
OBJECTIVE
To clone and express Schistosoma japonicum (Sj) calcyphosine gene, and purify the expressed protein.


METHODS
The encoding sequence selected from Sj cDNA library was amplified by PCR. After subcloned into prokaryotic expression vector pET-28a, the expressed protein was purified with His -Tag affinity chromatography. Western blotting was used to detect the immunogenicity. The structure and functions of the protein were analyzed by bioinformatics method, and the phylogenetic tree of the protein was drawn.


RESULTS
The recombinant protein was specifically recognized by the Sj infected rabbit serum. The bioinformatics analysis showed 4 EF-hand domains. Besides, it was predicted that Sj calcyphosine contains two phosphorylation sites for protein kinase C, eight phosphorylation sites for casein kinase II and one N-myristoylation site. The Sj calcyphosine belonged to type-II calcyphosine.


CONCLUSIONS
The calcyphosine gene is a calcium-binding protein and might be a potential candidate for diagnosis, vaccine or drug target.
Ju
Chuan
C
National Institute of Parasite Diseases, Chinese Centre for Disease Control and Prevention, Collaborating Center for Malaria, Schistosomiastis and Filariasis, Shanghai 200025, China.
Peng
Jian-xin
JX
Xu
Bin
B
Wang
Wei
W
Feng
Zheng
Z
Hu
Wei
W
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002135:Calcium-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D015801:Helminth Proteins / Q000235:genetics* / Q000378:metabolism; D007106:Immune Sera / Q000276:immunology; D008969:Molecular Sequence Data; D010802:Phylogeny; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D012549:Schistosoma japonicum / Q000235:genetics* / Q000276:immunology; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D002135:Calcium-Binding Proteins; D015801:Helminth Proteins; D007106:Immune Sera; D011994:Recombinant Proteins