兔豆状囊尾蚴TpRS1基因的克隆及其序列分析

摘要

旨在研究兔豆状囊尾蚴TpRS1的功能，根据GenBank中猪带绦虫RS1 cDNA序列设计引物，以豆状囊尾蚴总RNA为模板，利用RT-PCR技术首次克隆到兔豆状囊尾蚴TpRS1的完整开放阅读框序列，并利用生物信息学软件对该基因及其编码蛋白的基本理化性质、疏水性、信号肽、二级结构和抗原位点等方面进行了预测和分析。结果表明，兔豆状囊尾蚴TpRS1的cDNA序列为275 bp的基因片段，该序列含有一个由258个核苷酸组成的开放阅读框，编码85个氨基酸。其编码蛋白疏水性，分子质量为9.6 kD，理论等电点为9.07。TpRS1二级结构中α螺旋占70.59%，β折叠占5.88%，其余23.53%为无规卷曲，TpRS1没有信号肽序列，同源性分析表明与其他带科绦虫的一致性在72%以上，系统进化分析表明与泡状带绦虫处于同一进化分支。%To study the function of TpRS1 gene from Cysticercus pisiformis, the open reading frame(ORF)cDNA sequence of TpRS1 gene was cloned by RT-PCR from C. pisiformis total RNA with primers derived from Taenia solium RS1 gene sequence in the GenBank database. Biochemical properties, secondary structure, signal, hydrophobicity and antigenicity in TpRS1 protein were predicted by bioinformatics tools. The results indicated that TpRS1 cDNA sequence contained an ORF of 258 nucleotides and the deduced protein consisted of 85 amino acids with the theoretical molecular weight of 9.6 kD and isoelectric point of 9.07. Analysis of secondary structure revealed 70.59%and 5.88%ofα-helix andβ-strands, respectively, and others were loop. The signal peptide sequence of TpRS1 were not identified. TpRS1 protein sequence showed more than 72%identity with other Taenia spp.. Phylogenetic analysis indicated that TpRS1 located in the same clade with Taenia hydatigena.