骆驼斯氏副柔线虫NCC基因原核表达载体的构建及生物信息学分析

Bioinformatics Analysis and Construction of Prokaryotic Expression Vector of NCC Gene of Parabronema skrjabinin Camel

本试验旨在构建原核表达载体pET30a(+)-NCC,对斯氏副柔线虫NCC基因特性进行生物信息学分析。应用RT-PCR技术扩增斯氏副柔线虫NCC基因,将其克隆至原核表达载体pET-30a(+),利用生物信息学软件对序列结构与抗原表位进行分析。结果显示,NCC基因全长711bp,编码237个氨基酸,蛋白质理论大小值为25.252ku,理论等电点为6.33,蛋白质不稳定指数为40.08;跨膜结构和信号肽分析表明NCC蛋白存在1个跨膜区,无信号肽区域,其磷酸化位点分布位于丝氨酸(Ser)、苏氨酸(Thr)和酪氨酸(Tyr)残基上。NCC蛋白是一种抗原性较高的亲水性蛋白,既含有较多潜在的B细胞抗原表位又存在较多的T细胞抗原表位,本试验为斯氏副柔线虫诊断方法的建立及免疫防控提供参考。

In order to construct prokaryotic expression vector pET30a（＋）-NCC, the NCC gene from Para- bronema skrjabini was analyzed by bioinformatics.RT-PCR was used to obtain NCC gene,and the gene was cloned into the prokaryotic expression vector.The gene structure and epitopes were analyze by bioinformation software.The results indicated that NCC gene was 711 bp in length,encoding 237 amino acids,and the protein weights is 25.252 ku.Theoretical pI is 6.33 ,and instability index is 40.08.Transmembrane structure and signal peptide analysis showed that NCC protein has 1 transmembrane domain and no signal peptide region.The phosporylation site located in serine,threonine and tyrsine residues.NCC protein is a hydrophilic protein with high antigenicity. It contains more potential B cell antigen epitopes and more T cell antigen epitopes.This research provides a reference for the establishment of diagnostic methods and immunological control of Parabronerna skrjabini.