利用蛋白质组技术快速鉴定噬菌体结构蛋白

Rapid identification of phage structural proteins by proteomic techniques

为快速鉴定噬菌体的结构蛋白,并且发现具有新功能的蛋白,为噬菌体功能的研究提供基础。本研究用临床分离到的屎肠球菌为指示菌,从医院污水中分离到1株裂解性噬菌体,命名为v B_Efa P_IME199。对噬菌体大量培养、浓缩和纯化,然后进行蛋白质组学分析、全基因组测序,进化分析,确定该噬菌体为短尾噬菌体科,全基因组序列长为18839bp,GC含量为34.6%。在线注释预测该噬菌体基因组功能基因,注释出22个ORFs,其中11个ORFs为假想蛋白序列,其余11个ORFs的功能已知。采用质谱技术对浓缩纯化的噬菌体进行蛋白质组分析,共鉴定出11个蛋白质,其中有6个蛋白为功能已知的结构蛋白,其余为假想蛋白。本实验利用蛋白质组技术快速有效的鉴定出噬菌体的结构蛋白,建立了一种噬菌体蛋白的分析方法,为该噬菌体的基因功能研究提供了基础。

In order to rapidly identify phage structural proteins and to discover proteins with novel functions.It provided a basis for the study of phage functions. The bacterium used in this study was Enterococcus faecalis 1007, which was isolated from clinics, and a phage named vB_EfaP_IME 199, which was isolated from hospital sewage with Enterococcusfaecalis strain 1007 as the host. The phage was subjected to genome sequencing, phylogenetic tree analysis, proteomics analysis, and genome annotation. IME199 belongs to podoviridae. High throughput sequencing results showed the genome of IME199 was 18,839bp circular dsDNA, and had a G＋C content of 34.6%. The results of the gene annotation showed that there were 22 open reading frames （ORFs）. Of these 22 ORFs, 11 of them were hypothetical protein sequences, and the functions of the remaining 11 ORFs were known. Six proteins with known functions were identified as structure proteins of the phage particles by proteomics analysis. In this study, the proteome technology was used to identify the phage structure proteins quickly and effectively, and a phage protein analysis method was established. This study provides a basis for further functional researches of the structure genes.