[Cloning and sequence analysis of a novel stage-specific cDNA from adult Trichinella spiralis]
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Updated: Aug 30, 2026
Novel Sequence Discovery by Subtractive Genomics
Published on: January 25, 2019
Bao-quan Fu1, Feng Wang, Xiu-ping Wu
1Quartermaster University of PLA, Changchun 130062.
Objective:
To clone a stage-specific novel cDNA from 5 day-old adult worm (AD5) of Trichinella spiralis.
Methods:
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.
Results:
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%.
Conclusion:
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.
