Cloning and sequence analysis of a novel stage-specific cDNA from adult Trichinella spiralis

• Published in: Zhongguo jishengchongxue yu jishengchongping zazhi Vol. 21; no. 3; p. 154
• Main Authors: Fu, Bao-quan, Wang, Feng, Wu, Xiu-ping, Niu, Ting-xian, Lu, Qiang, Liu, Ming-yuan, Boireau, Pascal
• Format: Journal Article
• Language: Chinese
• Published: China 2003
• Subjects: Amino Acid Sequence; Animals; Base Sequence; Cloning, Molecular; DNA, Complementary - chemistry; DNA, Complementary - genetics; Genes, Helminth; Molecular Sequence Data; Open Reading Frames - genetics; Rats; Rats, Wistar; Swine; Trichinella spiralis - genetics
• ISSN: 1000-7423

To clone a stage-specific novel cDNA from 5 day-old adult worm (AD5) of Trichinella spiralis. The cDNA library of AD5 was screened by an AD5 stage-specific cDNA probe labeled with digoxigenin (DIG). The positive clones were sequenced and analysed. The positive clone contained a cDNA insert of 1,132 bp in length with a full length open reading frame (ORF) of 1,032 bp. The cDNA encoded a polypeptide of 343 amino acid residues(aa) with a molecular weight of 35.1 kDa and an isoelectric point (IP) of 4.8. InterProScan analysis showed that the 117-120 aa (SGYG) was a glycosaminoglycan attachment site, 27-86 aa was nematode cuticle collagen N-terminal domain and 153-228 aa was collagen repeat (G-x-y) domain. Signal PV2.0 analysis indicated that the region of 1-43 aa was a signal peptide. Blastn homology analysis in Genbank revealed that the cDNA had no obvious homology to any other known gene sequence. Blastn analysis revealed high homology to cuticle collagen with identities more than 40%. A novel AD5 stage-specific cDNA encoding a full length ORF was cloned and sequence analysis showed this gene encoded cuticle collagen of Trichinella spiralis.