论文部分内容阅读
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(open reading frame,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AD5 cDNA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(isoelectric point,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。Signal PV2.0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。
Objective To obtain cD-NA molecules encoding the open reading frame (ORF) of 5-day-old adults (AD5) of Trichinella spiralis. Methods AD5-specific cDNA fragment labeled with digoxigenin T671 was used as a probe to screen the AD5 cDNA library for nucleic acid hybridization. The positive clones screened were sequenced and analyzed by molecular biology software. As a result, a full-length 1132bp cDNA molecule was obtained. The cD-NA contained a complete ORF of 1032bp. The ORF encoded a polypeptide consisting of 343 amino acid residues. The deduced molecular weight was 35.1kDa and the isoelectric point isoelectric point, IP) was 4.8. InterProScan analysis showed that 117 to 120 amino acid residues (SGYG) were mucopolysaccharide binding sites, 27 to 86 amino acid residues were a nematode epidermal collagen N-terminal domain, and 153 to 328 amino acid residues were collagen helical triplets Repeat region (Gxy) domain. Signal PV2.0 analysis showed that the signal peptide in the region of 1 to 43 amino acids. Blastn homology analysis showed that there is no obvious homology with other known biological gene sequences and a new cDNA molecule. But higher homology with epidermal collagen, more than 40%. Conclusion A new cDNA molecule encoding the complete ORF encoding AD5-specific genes was obtained.