铜绿假单胞菌噬菌体PaP1主要结构蛋白编码基因的确定

Identification of the major structural protein coding gene of pseudomonas aerugmosa phage PaP1

目的:确定铜绿假单胞菌噬菌体PaP1主要结构蛋白的编码基因。方法:CsCl梯度离心纯化噬菌体PaP1 颗粒,通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离该噬菌体的结构蛋白,转印PVDF膜后,对主要结构蛋白进行N-端测序,根据测序结果和实测分子量,在PaP1基因组序列中检索推定基因表达产物分子量相近、 N-端氨基酸残基序列相同的蛋白质所对应的编码基因。结果:SDS-PAGE显示PaP1至少含有7种结构蛋白,其中相对分子质量为38400的蛋白是主要的结构蛋白,其N-端5个氨基酸残基顺序为:Ala-Asn-Thr-Arg-Ser。结论:噬菌体PaP1至少含有7个结构蛋白,其中相对分子质量为38 400的蛋白系主要结构蛋白,其编码基因为 ORF6841-7875。

Objective ： To identify the coding gene of the major structural protein of pseudomonas aeruginosa phage PAPl. Methods： Phage particles of PaPl were purified with the CsCl gradient centrifugation. After separated by SDS-PAGE, the phage structural proteins were transferred to a PVDF film, then the major band was excised from the film and subjected to N-terminal sequencing. The coding gene of the main structural protein could be identified by comparing its N-terminal sequences with amino acids sequences of the coding sequences of PaPl genome. Results ： PaPl particles are separated to at least seven bands by SDS-PAGE. The N-terminal sequences of the main structural protein are ： Ala-Asn-Thr-Arg-Ser. Conclusion： There are at least seven structural proteins in PaPl particles. The main structural protein is encoded by ORF6841-7875.