摘要
[目的]克隆沙眼衣原体(Chlamydia trachomatis)LpxA基因并分析其生物学特性。[方法]根据Ct LpxA基因序列设计特异性引物,利用PCR扩增LpxA基因,并把LpxA基因连接到p MD18-T载体,挑取阳性克隆进行PCR和DNA测序验证。最后使用生物信息学软件分析LpxA蛋白的生物学性质。[结果]从Ct基因组中PCR扩增得到840 bp的Ct LpxA基因,该基因共编码280个氨基酸。LpxA蛋白无信号肽,定位在细菌细胞质。二级结构预测得知α-螺旋占19.6%,延伸链占32.8%,β-转角为11.4%,无规卷曲为36%。三级结构预测得知3个相同的LpxA分子组成同源三聚体。预测得知LpxA蛋白有11个B细胞表位。[结论]克隆得到Ct LpxA基因,并预测了其结构和功能。
[Objective]To clone the LpxA gene from Chlamydia trachomatis and analyze its biological characters. [Methods]The specific primers were designed according the sequences of LpxA gene from C. trachomatis,which were used to amplify the target gene by using PCR. The LpxA gene was ligate to p MD18-T vector to create the recombinant which were confirmed by PCR and DNA sequencing. The biological characters of LpxA were further analyzed by using bioinformatics tools. [Results]The 840 bp length LpxA gene was cloned which encoding the LpxA protein. The LpxA protein was predicted to locate in the cytoplasm of the C. trachomatis and did not have signal peptides. The secondary structures of LpxA protein were predicted which showed its α-helix was 19. 6%,the extended strand was 32. 8%,the β turn was 11. 4% and the random coil was 36%. The tertiary structure of LpxA protein was predicted by homology modeling,which indicated it is a trimer of identical subunits. The LpxA protein has 11 B cell epitopes. [Conclusion] The Ct LpxA gene was cloned and its biological characters were analyzed.
出处
《生物技术》
CAS
2018年第1期44-48,共5页
Biotechnology
基金
特殊病原体防控湖南省重点实验室资助项目(湘科计字[2014]5号)
