Molecular cloning and characterisation of a putative aspartate proteinase associated with a gut membrane protein complex from adult Haemonchus contortus
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- 作者：Longbottom ; David ; Redmond ; Diane L. ; Russell ; Mary ; Liddell ; Susan ; Smith ; W. David ; Knox ; David P.
- 关键词：Haemonchus contortus ; Aspartyl proteinase ; Pepsinogen ; H-gal-GP ; Gut expression ; H-gal-GP ; Haemonchus galactose-containing glycoprotein ; Pep ; pepsinogen ; RT-PCR ; reverse transcribed polymerase chain reaction ; SL ; spliced leader ; IPTG ; isopropylthio-β
- 刊名：Molecular and Biochemical Parasitology
- 出版年：1997
- 出版时间：September, 1997
- 年：1997
- 卷：88
- 期：1-2
- 页码：63-72
- 全文大小：960 K
文摘
A cDNA was isolated from an adult Haemonchus contortus cDNA expression library the deduced amino acid sequence of which showed significant homology to mammalian pepsinogen sequences. The library was screened with antisera raised against Haemonchus galactose-containing glycoprotein complex, a gut membrane protein complex with aspartyl proteinase activity which has shown considerable potential as a protective antigen. The amino acid sequence obtained corresponded very closely in part to the N-terminal amino acid sequences of two polypetides within the complex. The enzyme was shown to be almost exclusively expressed by the blood-feeding parasite stages. The cDNA was expressed in E. coli, and antibody produced to the recombinant protein bound to the luminal surface of the gut in the adult parasite. The proteinase may play a central role in digesting the blood meal and is considered a potential sub-unit vaccine candidate.