[旋毛虫成虫期特异性新cDNA的克隆与序列分析]

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

作者信息

Fu Bao-quan, Wang Feng, Wu Xiu-ping, Niu Ting-xian, Lu Qiang, Liu Ming-yuan, Boireau Pascal

机构信息

Quartermaster University of PLA, Changchun 130062.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2003;21(3):154-6.

PMID:14628346

Abstract

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.

摘要

目的

从旋毛虫5日龄成虫（AD5）中克隆一个阶段特异性新cDNA。

方法

用地高辛（DIG）标记的AD5阶段特异性cDNA探针筛选AD5的cDNA文库。对阳性克隆进行测序和分析。

结果

阳性克隆包含一个长度为1132 bp的cDNA插入片段，其全长开放阅读框（ORF）为1032 bp。该cDNA编码一个由343个氨基酸残基组成的多肽，分子量为35.1 kDa，等电点（IP）为4.8。InterProScan分析显示，第117 - 120个氨基酸（SGYG）为糖胺聚糖附着位点，第27 - 86个氨基酸为线虫表皮胶原蛋白N端结构域，第153 - 228个氨基酸为胶原蛋白重复（G - x - y）结构域。Signal PV2.0分析表明，第1 - 43个氨基酸区域为信号肽。在Genbank中进行Blastn同源性分析显示，该cDNA与任何其他已知基因序列均无明显同源性。Blastn分析显示与表皮胶原蛋白具有高度同源性，一致性超过40%。