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| 旋毛虫成虫期特异性基因全长cDNA的克隆与序列分析 | |
| 引用本文： | 付宝权,王峰,吴秀萍,牛廷献,卢强,刘明远,Pascal Boireau. 旋毛虫成虫期特异性基因全长cDNA的克隆与序列分析[J]. 中国寄生虫学与寄生虫病杂志, 2003, 21(3): 154-156 |
| 作者姓名： | 付宝权 王峰 吴秀萍 牛廷献 卢强 刘明远 Pascal Boireau |
| 作者单位： | 1. 解放军军需大学,长春,130062 2. 兰州军区兰州总医院,兰州,730050 3. 法国食品卫生安全局,巴黎,94703 |
| 基金项目： | 国家自然科学资助项目(NSFC0170709)，中法先进研究计划(BT96.12)资助项目 |
| 摘 要： | 目的获取编码旋毛虫5日龄成虫(AD5)期特异性基因完整开放阅读框架(openreadingframe,ORF)cD-NA分子。方法利用地高辛标记的AD5期特异性cDNA片段T671作为探针对AD5cDNA文库进行核酸杂交筛选,将筛选出的阳性克隆进行测序,利用分子生物学软件进行分析。结果获得1个全长1132bp的cDNA分子,该cD-NA含有1个1032bp完整的ORF,该ORF编码1个343个氨基酸残基组成的多肽,其分子量理论推导值为35.1kDa,等电点(isoelectricpoint,IP)为4.8。InterProScan分析显示117～120氨基酸残基(SGYG)为粘多糖结合位点,27～86氨基酸残基为一线虫表皮胶原蛋白N-末端区结构域,153～328氨基酸残基为胶原蛋白螺旋三联体重复区(G-x-y)结构域。SignalPV2.0分析显示,在1～43氨基酸区域为信号肽。Blastn同源性分析表明,与其它已知生物基因序列无明显的同源性,为一新的cDNA分子。但与表皮胶原蛋白同源性较高,大于40％。结论筛选获得一个新的编码AD5期特异性基因的完整ORF的cDNA分子。 |
| 关 键 词： | 旋毛虫 5日龄成虫期特异性基因 cDNA克隆 序列分析 |
| 文章编号： | 1000-7423(2003)-03-0154-03 |
| 修稿时间： | 2003-06-24 |
Cloning and Sequence Analysis of a Novel Stage-Specific cDNA from Adult Trichinella spiralis |
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| FU Bao-quan,WANG Feng,WU Xiu-ping,NIU Ting-xian,LU Qiang,LIU Ming-yuan,Pascal Boireau. Cloning and Sequence Analysis of a Novel Stage-Specific cDNA from Adult Trichinella spiralis[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2003, 21(3): 154-156 | |
| Authors: | FU Bao-quan WANG Feng WU Xiu-ping NIU Ting-xian LU Qiang LIU Ming-yuan Pascal Boireau |
| Affiliation: | Quartermaster University of PLA, Changchun 130062. |
| Abstract: | Objective To clone a stage-specific novel cDNA from 5 day-old adult worm (ADS) 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 singal peptide. Blastn homology analysis in Genbank revealed that the cDNA had no obvious homology to any other known gene sequence. Blastp analysis revealed high homology to cuticle collagen with identities more than 40 % . Conclusion A novel ADS stage-specific cDNA encoding a full length ORF was cloned and sequence analysis showed this gene encoded cuticle collagen of Trichinella spiralis. |
| Keywords: | Trichinella spiralis stage-specific gene cDNA cloning sequence analysis |
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