Cloning, characterization and DNA immunization of an Onchocerca volvulus glyceraldehyde-3-phosphate dehydrogenase (Ov-GAPDH)

被引:32
作者
Erttmann, KD
Kleensang, A
Schneider, E
Hammerschmidt, S
Büttner, DW
Gallin, M
机构
[1] Bernhard Nocht Inst Trop Med, D-20359 Hamburg, Germany
[2] Univ Wurzburg, Res Ctr Infect Dis, D-97070 Wurzburg, Germany
来源
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR BASIS OF DISEASE | 2005年 / 1741卷 / 1-2期
关键词
glyceraldehydc-3-phosphate dehydrogenase; filaria; Onchocerca volvulus; plasminogen-binding; protective antigen; DNA immunization;
D O I
10.1016/j.bbadis.2004.12.010
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
in the search for Onchocerca volvulus antigens possibly involved in protection against human onchocerciasis, partial amino acid sequence analysis of one of the O. volvulus antigens of the serologically identified proteins showed a close relationship to the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein family. Subsequent adult worm cDNA library screening and cloning produced a clone of 1650 bp. An open reading frame spans over 1020 bp encoding for a protein of 340 amino acids with an apparent molecular weight of 38000. Comparison of the complete amino acid sequence identified this protein as a member of the GAPDH protein family. The recombinantly expressed protein shows GAPDH enzymatic activity as well as plasminogen-binding capacity. DNA sequence analysis of the corresponding gene revealed the presence of two introns. Using immunohistology Ov-GAPDH Was observed in microfilariae, infective larvae, and adult male and female worms. Most striking was the labelling of the musculature of the body wall. Labelling was also observed in the pseudocoeloma cavity and in a subset of cell nuclei, suggesting additional, non-glycolytic functions of the Ov-GAPDH. Gene gun immunization with the DNA-construct in cattle led to specific humoral immune responses. Thus, the protective potential of the DNA-construct of Ov-GAPDH can be evaluated in vaccination trials using animal models such as the cattle/Onchocerea ochengi model. (c) 2004 Elsevier B.V. All rights reserved.
引用
收藏
页码:85 / 94
页数:10
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