[Cloning and expression of gynecophoral canal protein gene of Schistosoma japonicum (Chinese strain)].
A 1949 bp cDNA fragment was amplified by RT-PCR from adult Schistosoma japonicum (Chinese strain) mRNA with 3 pair of primers that were designed according to published SmGCP gene encoding gynecophoral canal protein of Schistosoma mansoni and SjGCP1 gene encoding the conservative region of gynecophoral canal protein of Schistosoma japonicum. Sequence analysis indicated that this fragment, named SjGCP, with 85% identity to SmGCP, contained a complete open reading frame (ORF) of gynecophoral canal protein gene of Schistosoma japonicum (Chinese strain). The amino acid sequence shared 83.7% identity with gynecophoral canal protein of Schistosoma mansoni. This fragment was cloned into the expression vector pET28c(+) and subsequently expressed in Escherichia coli. SDS-PAGE revealed that the molecular weight of this expressed product was 80 kD. Western blotting showed that the recombinant protein reacted well with the rabbit serum immunized with Sj worm antigen, indicating that this expressed product had good antigenicity.
Jin
Y M
YM
Shanghai Institute of Animal Parasitology, Chinese Academy of Agricultural Science, Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai 200232, China.
Lin
J J
JJ
Feng
X G
XG
Zhang
L
L
Wu
X F
XF
Zhou
Y C
YC
Cai
Y M
YM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis; D018509:DNA, Helminth / Q000032:analysis; D004926:Escherichia coli; D015870:Gene Expression; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D015801:Helminth Proteins / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D012549:Schistosoma japonicum / Q000235:genetics* / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D018509:DNA, Helminth; D006023:Glycoproteins; D015801:Helminth Proteins; C103128:gynecophoral canal protein, 86-kDa