溶藻弧菌去乙酰化酶基因cobB克隆及其功能验证

Molecular Cloning and Functional Verification of Histone Deacetylase Gene cobB in Vibrio alginolyticus

旨在研究溶藻弧菌(Vibrio alginolyticus)去乙酰化酶cobB基因的功能。进行了cobB基因的克隆,生物信息学分析,CobB蛋白质诱导表达、纯化及功能研究。CobB蛋白质的相对分子质量为27.09750 kD,理论等电点为5.15,化学式为C_(1186)H_(1865)N_(339)O_(369)S_(10),亲水性平均系数(GRAVY):-0.409,为亲水蛋白。CobB与同弧菌属的魔鬼弧菌(Vibrio diabolicus),哈维弧菌(Vibrio harveyi),霍乱弧菌(Vibrio cholerae)和副溶血性弧菌(Vibrio parahaemolyticus)具有高度同源性,在革兰氏阴性菌中相对保守;融合蛋白大小约为53 kD,去除标签后CobB蛋白分子量约为27 kD;CobB蛋白最适表达条件为37℃,0.4 mmol /L IPTG下诱导4 h。功能分析表明,CobB对乙酰化蛋白具有去乙酰化作用。

This work aims to study the function of the histone deacetylase cobB gene in Vibrio alginolyticus.The clone of cobB gene,its bioinformatics analysis,and induction,expression,purification and function of CobB protein were carried out.The relative molecular mass was 27.09750 kD,theoretical isoelectric point was 5.15,the chemical formula of the protein was C_(1186)H_(1865)N_(339)O_(369)S_(10),and hydrophilic average coefficient grand aural of hydropathity(GRAVY):-0.409,showing that it was a hydrophilic protein.CobB presented high homology with that of Vibrio diabolicus,Vibrio harveyi,Vibrio cholerae,and Vibrio parahaemolyticus,and was relatively conserved among Gram-negative bacteria.The fusion protein was about 53 kD,and the molecular weight of the CobB protein after tag removed was about 27 kD.The optimal expression condition of the CobB protein was 37℃ and inducing time 4 h with 0.4 mmol/L IPTG.Functional analysis showed that the CobB had deacetylation effect on acetylated protein.