[Cloning and expression of 21.7 kD protein gene of Schistosoma japonicum (Chinese strain)].
A 558 bp cDNA fragment was amplified by RT-PCR from adult Schistosoma japonicum(Chinese strain) mRNA with a pair of primers that were designed according to published Sj21.7p gene encoding 21.7 kD protein of Schistosoma japonicum(Philippines strain). Sequence analysis indicated that this frame, named Sj21.7 (Ch), with 99% homology to Sj21.7 p, contained a complete open reading fragment (ORF) of 21.7 kD protein gene of Schistosoma japonicum(Chinese strain). The amino acid sequence shared 98% homology with 21.7 kD protein of Schistosoma japonicum. This fragment was cloned into the expression vector pET28a (+) and subsequently expressed in Escherichia coli with IPTG induction. SDS-PAGE analysis revealed that the molecular weight of this expressed product was 25.4 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
Ya-Mei
YM
Shanghai Institute of Animal Parasitology, Chinese Academy of Agricultural Science, Key Laboratory of Animal Parasitology, Ministry of Agriculture of China, Shanghai Institute for Biological Sciences, Shanghai, China.
Lin
Jiao-Jiao
JJ
Zhang
Liang
L
Ni
Zhen-Ya
ZY
Fu
Zhi-Qiang
ZQ
Wu
Xiang-Fu
XF
Zhou
Yuan-Cong
YC
Cai
You-Min
YM
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D015801:Helminth Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D011817:Rabbits; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000276:immunology; D012549:Schistosoma japonicum / Q000235:genetics*; D017385:Sequence Homology
D015801:Helminth Proteins; D011994:Recombinant Proteins