奶牛乳腺炎无乳链球菌菌毛岛屿PI-2a骨架蛋白BP基因的克隆及其序列分析

发布时间：2018-11-02 18:50

【摘要】：目的获得通辽地区奶牛乳腺炎致病菌无乳链球菌相关菌毛岛屿BP基因序列并进行比对分析。方法根据DNAMAN分析基因库中牛源无乳链球菌菌毛岛屿PI-2a的BP基因序列设计1对特异性引物,以无乳链球菌通辽分离株总DNA为模板,PCR扩增BP基因片段并进行序列比对分析。结果获得了菌毛岛屿PI-2a的BP基因序列,目的基因为609bp,编码203个氨基酸,其基因序列与GenBank上公布的牛源无乳链球菌菌毛岛屿PI-2a的BP基因(EU930008)同源性为100%。BP蛋白二级结构的α螺旋在N端分布较多,β折叠较丰富且分布C端较多,无规则卷曲分布较均匀。BP蛋白的亲水区分布较广,遍布于整个蛋白区,属于亲水性蛋白。BP蛋白分子表面可及性较高,区域占比达57.0%,且具有抗原性。结论获得了奶牛乳腺炎无乳链球菌通辽分离株菌毛岛屿PI-2a骨架蛋白BP基因序列,该基因序列具有高度保守性,其编码蛋白具有抗原表位,可作为奶牛无乳链球菌重组基因工程疫苗候选亚单位抗原。
[Abstract]:Objective to obtain the BP gene sequence of Streptococcus lactis related island in Tongliao area. Methods A pair of specific primers were designed according to the BP gene sequence of PI-2a from Streptococcus lactobacillus pili from DNAMAN gene bank. The total DNA of Streptococcus lactis Tongliao isolate was used as template, and the BP gene fragment was amplified by PCR and compared with the sequence analysis. Results the BP gene sequence of the pili island PI-2a was obtained. The target gene was 609 BP encoding 203 amino acids. The BP gene (EU930008) homologous to the BP gene (EU930008) of Streptococcus lactococcus pili PI-2a published on GenBank was found to be a 伪 helix with a secondary structure of 100%.BP protein. The 伪 -helix of 100%.BP protein was found to be more abundant in N terminal, rich in 尾 folding and more in C-terminal distribution. The hydrophilic region of BP protein was widely distributed in the whole protein region and belonged to hydrophilic protein. The surface accessibility of BP protein was high, and the area of Prida protein was 57.0 and antigenicity. Conclusion the sequence of PI-2a skeleton protein BP gene in the tuber island of Streptococcus lactis is highly conserved and its coding protein has antigenic epitopes. It can be used as candidate subunit antigen for recombinant genetic engineering vaccine of streptococcus acuminata in dairy cattle.
【作者单位】：内蒙古民族大学生命科学学院内蒙古自治区乳源性致病菌防控工程技术研究中心内蒙古民族大学乳源性致病菌研究所;
【基金】：国家自然科学基金项目(No.31560689) 内蒙古自治区乳源性致病菌防控工程技术研究中心开放课题(No.MDK2016036)
【分类号】：S852.61

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