[Gene cloning and characterization of mitochondria-related protein of Schistosoma japonicum].
OBJECTIVE
To subclone and characterize a cDNA clone coding for Schistosoma japonicum (S.j.) mitochondria-related protein.


METHODS
The open reading frame of the fragment(Sj338/24) obtained from an adult worm cDNA library of S.j. was analysed, at the upstream and downstream of the open reading frame(ORF) the primers A and B were designed, respectively, and the cDNA fragment was used as PCR template. The Sj338 gene fragment obtained was amplified by PCR method and then subcloned into pGEM-T vector for sequencing. The gene sequence was analyzed and the target fragment was restrictedly digested and subcloned into expression vector pGEX-6P-1. The expressed recombinant protein was purified and characterized.


RESULTS
The cloned Sj338 gene was demonstrated to be 487 bp long containing one 459 bp ORF, encoding a protein with a molecular weight of 17 kDa. The nucleotide sequence of the cloned gene Sj338 had higher homology with those genes coding for mitochondrial outer membrane protein of Homo sapiens and Rattus norvegicus. The recombinant construct of pGEX-6P-1/Sj338 could be expressed efficiently and the antigenicity of its product rSj338 has been demonstrated by Western blotting.


CONCLUSIONS
Sj338 may be the gene coding for S.j. mitochondria-related protein and the recombinant protein may be used as a new vaccine candidate.
Hu
X M
XM
Department of Parasitology, Binzhou Medical College, Binzhou 256603.
Wu
H W
HW
Zhang
Z S
ZS
Su
C
C
Zhao
W
W
Shen
L
L
Wang
R Z
RZ
Ma
L
L
Zhou
J L
JL
Chen
S Z
SZ
Wu
G L
GL
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015801:Helminth Proteins / Q000096:biosynthesis / Q000235:genetics*; D008928:Mitochondria; D008969:Molecular Sequence Data; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D012549:Schistosoma japonicum / Q000235:genetics*
D018076:DNA, Complementary; D015801:Helminth Proteins; D011994:Recombinant Proteins