cDNA cloning and location of Pagumogonimus skrj abini cysteine protease

• Published in: Zhongguo jishengchongxue yu jishengchongping zazhi Vol. 21; no. 4; p. 214
• Main Authors: Wang, Ying, Zhang, Xi-lin, Zhang, Yan-ling, Duan, Jian-hua, Zhang, Jing-ru, Huang, Fu-sheng
• Format: Journal Article
• Language: Chinese
• Published: China 2003
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Cloning, Molecular; Cysteine Endopeptidases - biosynthesis; Cysteine Endopeptidases - genetics; DNA, Complementary - genetics; Gene Expression; In Situ Hybridization; Molecular Sequence Data; Paragonimus - enzymology; Paragonimus - genetics; Reverse Transcriptase Polymerase Chain Reaction
• ISSN: 1000-7423

To clone the cysteine protease cDNA fragment from Pagumogonimus skrjabini adults and locate the tissue of the adult worm where cysteine protease is expressed. The cysteine protease cDNA fragment was amplified by reverse transcription-polymerase chain reaction (RT-PCR) with degenerated primers. The production was TA-cloned into the pUCm-T vector and sequenced. DNASIS program was used to analyse the nucleotide sequence and deduce the amino acid sequence, which was aligned with the correlated parasite cysteine protease afterwards. The digoxin labeled cRNA probe was synthesised by in vitro transcription with the cloned cDNA as template. The frozen sections of the adult worms were analysed by hybridization in situ to locate the gene expression. A 495 bp cDNA fragment was amplified by RT-PCR and sequenced. An amino acid sequence was deduced by DNASIS. Sequence analysis and alignment showed significant homologies with the correlated parasite cysteine proteinases and conservation of Cys, His and Asn residues that fro