旋毛虫5日龄成虫期特异性基因的筛选与序列分析

目的　获得旋毛虫 5日龄成虫期特异性基因序列。方法　应用旋毛虫 5日龄成虫期待异性探针对 5日龄成虫cDNA文库进行杂交筛选、克隆测序及核苷酸序列数据库 (GenBank)查寻比较 ,登陆InterProScan进行基因、氨基酸序列分析。结果　利用消减杂交 (subtractedhybridization)获得的旋毛虫 5日龄成虫期特异性探针筛选 5日龄成虫cDNA文库 ,获得一个长 1 1 32bp的基因。其开放阅读框架由 1 0 30个核苷酸组成 ,编码 343个氨基酸。序列分析表明 ,该序列是一个尚未见报道的旋毛虫基因序列 ,在序列 1 1 7～ 1 2 0位氨基酸为氨基葡聚糖 (GAG)结合位点 ,在 2 7～ 86 ,1 5 3～ 32 8位氨基酸处存在Gly -X -Y胶原蛋白重复序列 ,分别编码两个胶原蛋白多肽。结论　获得旋毛虫 5日龄成虫期特异性基因 ,其编码产物为表皮胶原蛋白

Screening and Sequence Analysis of the 5 Days Old Adult Stage-specific Gene of Trichinella spirais

objective: To obtain the sequence of the 5 days old adult stage-specific gene of Trichinella spirais. Method: An t days old abult T.spirais cDNA library was screened using an 5 days old adult T.spirais stage-specific probe. The gene and amino acid sequence was analyzed by Landing GeneBank database and InterProScan. Results: A cDNA clone of 1132bp was obtained by screening the library with an 5 days old adult stage-specific probe. The clone contained one open reading frame composed of 1030bp encoding 343 amino acids. GenBank searching revealed that no identical cDNA was found. One Glycosaminoglycan attachment site located at the positions within amino acids117-120. Gly-X-Y cuticle collagen repeat sequence were predicted within the amino acids 27-86, 153-328, which encode two collagen-like polypeptides respectively. Conclusion: A novel stage-specific gene of 5 days old adult T.spirais encoding certain predicted cuticle collagen polypeptides was obtained.