[Cloning, expressing and characterizing of a phosphoglycerate mutase gene of Schistosoma japonncum].
Phosphoglycerate mutase (PGAM) is a key enzyme in glycolytic pathways. With PCR technique based on an EST identified in our lab, a novel gene named SjPGAM (GenBank Accession No. EU374631) was cloned. Sequence analysis revealed that the ORF of SjPGAM gene contained 753 nucleotides, encoding 250 amino acids, and the molecular weight was about 28.26 kD. Real-time PCR analysis showed that the mRNA level of SjPGAM was much higher in the 14 days and 19 days schistosomula than other stages, suggesting that the gene was a schistosomula stage differential expression gene. The SjPGAM cDNA fragment was subcloned into an expression vector pET-28a (+) and transformed into Escherichia coli BL21 cells. In the presence of IPTG, the 31 kD fusion protein was expressed in included bodies. Western blotting revealed that the fusion protein could be recognized by the rabbit serum anti-Schistosoma japonicum adult worm antigen preparation. The study provides important basis for investigating the mechanism of the PGAM in the glycolytic pathways of Schistosoma japonnicum.
Zhou
Yan
Y
National Laboratory of Animal Schistosomiasis Control, Key Laboratory of Animal Parasitology Ministry of Agriculture, Shanghai Veterinary Research Institute of CAAS, Shanghai 200232, China.
Lin
Jiaojiao
J
Yao
Lixiao
L
Wang
Xinzhi
X
Shi
Yaojun
Y
Lu
Ke
K
Liu
JinMing
J
Fu
Zhiqiang
Z
Tao
Lihong
L
D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D008297:Male; D010736:Phosphoglycerate Mutase / Q000235:genetics* / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism*; D012549:Schistosoma japonicum / Q000201:enzymology* / Q000235:genetics; D012554:Schistosomiasis japonica / Q000276:immunology / Q000469:parasitology
D012333:RNA, Messenger; D011994:Recombinant Proteins; D010736:Phosphoglycerate Mutase