衣原体噬菌体phiCPG1衣壳蛋白Vp1的空间结构分析及分段克隆表达  

The Spatial Structure Analysis and Segmental Cloning of the Chlamydiaphage phi CPG1 Capsid Protein Vp1

作　　者：周全[1] 刘原君[1] 孙长贵[1] 郭媛丽[1] 马璟玥[1] 郑蕾[1] 刘全忠[1] 

机构地区：[1]天津医科大学总医院皮肤性病科,天津300052

出　　处：《中国皮肤性病学杂志》2016年第5期441-444,共4页The Chinese Journal of Dermatovenereology

基　　金：国家自然科学基金面上项目(31370211)

摘　　要：目的预测衣原体噬菌体phi CPG1衣壳蛋白Vp1的空间结构,并将Vp1表达为更小的蛋白片段,为后续实验奠定基础。方法通过蛋白空间结构分析网站I-TASSER和Predict Protein对phi CPG1衣壳蛋白Vp1的空间结构进行预测,应用TA克隆的方法将Vp1分为不同的部分进行克隆表达,最后通过Western blot技术分别对目的蛋白进行鉴定。结果根据空间结构预测结果以及相关背景资料将衣原体噬菌体phi CPG1衣壳蛋白Vp1分为Vp11-189和Vp1190-502两部分进行克隆表达,目的蛋白Vp11-189和Vp1190-502的基因序列长度分别为567bp和939bp,经检索确定两蛋白碱基序列与Genebank结果相同。而且,Western blot结果显示两目的蛋白的相对分子质量分别为20k Da和35k Da。结论成功将衣原体噬菌体phi CPG1衣壳蛋白Vp1分为Vp11-189和Vp1190-502两部分进行表达,这为后期的深入研究提供了一定的实验基础。Objective To predict the spatial structure of the chlamydiaphage phiCPG1 capsid protein Vpl,and to produce smaller-sized protein in order to provide a basis for further studies. Methods The websites I-TASSER and PredictProtein were used to predict the spatial structure of the chlamydiaphage phiCPG1 capsid protein Vpl. Various fragments of chlamydiaphage phiCPG1 capsid protein Vpl were cloned with TA-cloning technique, and identified by Western blot. Results According to the predicted results and the relevant background information, Vp1 1-189 and Vp1 190-502 were cloned. The sequence sizes of Vp1 1-189 and Vp1190-502 genes were 567bp and 939bp, respectively,which base sequences were the same as that in the Genebank. The Western blot revealed that the relative molecular weight of Vp1 1-189 and Vp1 190-502 were 20kDa and 35kDa respectively. Conclusion Two fragments chlamydiaphage phiCPG1 capsid protein Vp1, Vp1 1-189 and Vp1 190-502, have been cloned, providing a valuable reference for future studies.

关 键 词：衣原体噬菌体 phiCPG1 VP1蛋白 

分 类 号：R374.1[医药卫生—病原生物学]