鸭疫里默氏杆菌Ⅸ型分泌系统分泌蛋白AS87_RS03090的原核表达及其突变株构建

Prokaryotic Expression and Mutant Construction of the As87_Rs03090,A Secretory Protein from the Riemerella anatipestifer TypeⅨSecretion System

研究发现细菌Ⅸ型分泌系统(T9SS)效应分子的生物学功能具有多样性,它们参与细菌自身的生命活动或与细菌的致病性密切相关。在前期研究中我们发现鸭疫里默氏杆菌T9SS的效应分子大多具有酶活性,可能参与其生命活动或与致病性相关。为探索鸭疫里默氏杆菌T9SS分泌蛋白AS87_RS03090的功能及其与细菌毒力的关系,本研究根据鸭疫里默氏杆菌AS87_RS03090基因序列设计了特异性引物,并在其上、下游引物中分别添加BamHⅠ和SalⅠ酶切位点,将其克隆到pCold TF载体上构建原核表达载体,诱导表达和纯化出鸭疫里默氏杆菌AS87_RS03090重组蛋白(rAS87_RS03090);本研究还利用自然转化的方式成功构建了AS87_RS03090基因突变株Yb2Δ03090。研究结果将有助于进一步鉴定鸭疫里默氏杆菌T9SS在其致病过程中发挥的作用。

It has been found that eff ector molecules of bacterial type IX secretion system(T9SS)play multiple functions on participating in the bacterial life activities or pathogenicity.In previous studies,we found that most T9SS eff ector molecules of Riemerella anatipestifer had enzymatic activity,which might be involved in its life activities or related to pathogenicity.In order to study the function of R.anatipestifer T9SS secreted protein AS87_RS03090 and its relationship with bacterial virulence,specifi c primers were designed in this study based on the gene sequence of AS87_RS03090 with induced Bam HⅠand SalⅠsites.Then pCold TF vector was constructed for expression of the recombinant protein AS87_RS03090.Moreover,the AS87_RS03090 mutant strain Yb2Δ03090 was also constructed by natural transformation.The results of this study were helpful to further study the role of R.anatipestifer T9SS in pathogenesis.