浅绿色气球菌噬菌体vB_AviM_AVP编码蛋白的表达纯化和功能分析

Expression,purification and functional analysis of the protein encoded by the bacteriophage vB_AviM_AVP of Aeroeoccus viridans

根据已报道的全球首个浅绿色气球菌噬菌体vB_AviM_AVP的基因组,通过比对分析对各个开放阅读框功能的预测,从中选取以下除裂解酶AVPL外可能具有裂解活性的蛋白基因ORF86、ORF90、ORF94、ORF98进行原核表达与纯化。试验表明,LysORF86、LysORF94和LysORF90可成功表达,但是仅在酶谱试验中对不同细菌表现出了活性;直接平板涂布滴板试验、利用高碘酸以及蛋白酶K处理菌体试验及混合蛋白活性检测试验中,均未表现出活性。本研究为全球首个浅绿色气球菌噬菌体vB_AviM_AVP的进一步研究奠定基础。

According to the genome of Aeroeoccus viridans bacteriophage vBAviMAVP reported firstly in the world,to predict the function of each open reading frame by comparison and analysis.In addition to Lyase AVPL,the ORF86,ORF90,ORF94 and ORF98 protein genes that may have cleavage activity were selected for prokaryotic expression and purification.The results showed that LysORF86,LysORF94 and LysORF90 were expressed successfully,but only showed activity against different bacteria in zymogram experiment,and showed no activity in the direct plate coating drop plate test,bacterial zymogram with periodate and proteinase K,and the mixed protein activity detection test.This study laid a theoretical and experimental foundation for the further study of Aeroeoccus viridans bacteriophage vBAviMAVP.