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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 capsid protein Vp1 of chlamydial phage phi CPG1 and to express Vp1 as a smaller protein fragment, which laid the foundation for further experiments. Methods The spatial structure of phi CPG1 capsid protein Vp1 was predicted by the protein structure analysis website I-TASSER and Predict Protein. The Vp1 was cloned and expressed in different fractions by TA cloning method. Finally, Protein identification. Results According to the results of spatial structure prediction and related background data, the capsid protein Vp1 of phage phi CPG1 was divided into two parts: Vp11-189 and Vp1190-502. The length of gene sequences of Vp11-189 and Vp1190-502 were 567 bp And 939bp. The two protein base sequences were found to be the same as Genebank results. Moreover, the results of Western blot showed that the molecular weights of the two proteins were 20kDa and 35kDa, respectively. Conclusions The VP1 capsid protein Vp1 of phage phage phi CPG1 was successfully expressed in two parts of Vp11-189 and Vp1190-502, which provided some experimental basis for further study.