CLONING, MOLECULAR CHARACTERIZATION, AND FUNCTIONAL-ACTIVITY OF SCHISTOSOMA-JAPONICUM GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE, A PUTATIVE VACCINE CANDIDATE AGAINST SCHISTOSOMIASIS-JAPONICA

被引：51

作者：

WAINE, GJ ^{[1
]}

BECKER, M ^{[1
]}

YANG, W ^{[1
]}

KALINNA, B ^{[1
]}

MCMANUS, DP ^{[1
]}

机构：

[1] QUEENSLAND INST MED RES,BANCROFT CTR,TROP HLTH PROGRAM,MOLEC PARASITOL UNIT,300 HERSTON RD,BRISBANE,QLD 4029,AUSTRALIA

来源：

INFECTION AND IMMUNITY | 1993年 / 61卷 / 11期

关键词：

D O I：

10.1128/IAI.61.11.4716-4723.1993

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

We report the cloning, molecular characterization, and purification of functionally active recombinant glyceraldehyde-3-phosphate dehydrogenase (GAPDH) from the human bloodfluke Schistosoma japonicum. The GAPDH homolog from the related species Schistosoma mansoni has shown correlation of antibody titer to resistance to reinfection. A 1,164-bp cDNA (C1) was isolated from an S. japonicum lambdaZapII cDNA expression library immunoscreened with hyperimmune rabbit serum raised against soluble adult S. japonicum proteins. The open reading frame of C1 encodes a protein of 338 amino acids exhibiting 90% identity to the amino acid sequence of S. mansoni GAPDH. The inferred molecular mass of the protein is 36,589 daltons, and in vitro translation of the cDNA With [S-35]methionine produced a radiolabelled band of the predicted size. Antibodies to C1 selected from hyperimmune rabbit serum by affinity purification recognized an S. japonicum protein doublet of 37 kDa but did not cross-react with a corresponding protein in S. mansoni extracts. The S. japonicum GAPDH appears to be translated from a single mRNA encoded by a single-copy gene. After subcloning in the QIAexpress vector pQE-10 and subsequent expression, the recombinant protein was purified under nondenaturing conditions and shown to exhibit functional GAPDH enzymatic activity.

引用

页码：4716 / 4723

页数：8

共 29 条

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