Erwinia herbicola冰核活性蛋白的分离、电泳分析鉴定

Differential centrifugal isolation and identification of electrophoresis analysis to ice-nucleating activity protein of Erwinia herbicola

对Erwiniaherbicola(A2 5 )菌株的冰核活性蛋白的分离纯化及电泳进行研究。主要方法①按双温培养的方法 ,获得了较高的生物量积累和强冰核活性的诱导表达。②实验采用渗透压冲击法破碎细菌细胞 ,破碎率达 98 6 7%。③通过差速离心法获取不同冰核活性蛋白组分 ,测定各组分的冰核活性和SDS -PAGE电泳图谱分析。建立了冰核活性因子的高冰核活性与离心力之间的关系 ;利用SDS -PAGE还建立了具冰核活性蛋白的分子量的大小与冰核活性蛋白组分之间的关系。证实了具高冰核活性蛋白质最小结构单位约为 2 6 0kD。

The ice-nucleation protein isolated and purified of Erwinia herbicola (A25) bacterial strain and the ice-nucleation proteins of electrophoresis characteristic is investigated in this paper Method:①According to the methods of the two-temperatures,the higher biomass accumulated and the highest ice-nucleation expressed by inducing is obtained ②The bacterial cell are osmotically shocked in this experiment,the rate of shatter come to 98 67% ③The differ fractions of the ice-nucleation activity proteins are collected by the differential centrifugation Results:The relations between the ice-nucleating activity of ice-nucleation proteins and centrifugal force are established by measuring the ice-nucleating activity of fractions and analysis of the SDS-PAGE electrophrasis The relationship between the molecular weight with the ice-nucleating activity proteins and the fractions with the ice-nucleating activity protein are also founded Conclusion:The least structural unit confirmed with higher ice-nucleating activity proteins(-5℃)is about 26 0kD