摘要
用扁豆几丁酶免疫家兔，获得抗扁豆几丁酶的抗体，将此抗体与Ｓｅｐｈａｒｏｓｅ ４Ｂ偶联，制备免疫亲和吸附剂，用以纯化苦瓜几丁酶．苦瓜叶片的粗提液经过免疫亲和吸附柱后，可获得电泳纯的几丁酶，其分子量为３５ ｋＤ，与用几丁质凝胶为亲和吸附剂的纯化结果一致．表明利用植物几丁酶在结构上的保守性，用免疫亲和法可纯化不同植物的同类几丁酶．与几丁质凝胶亲和柱相比，免疫亲和法纯化植物几丁酶具有快速、亲和柱可重复使用等的优点．利用免疫亲和层析获得的纯化样品，研究了苦瓜几丁酶对真菌的抑制试验，研究结果表明，苦瓜几丁酶能分解棉花枯萎病菌的菌丝体细胞壁制备物，并对其孢子芽管的伸长有一定抑制作用．
The imuno affinity absorbet gel that resulted from antiserum of rabbits against hyacinth pea pod chitinase coupled with Sepharose 4B was prepared for purification of chitinase in Momordica charantia leaves.By one step through the immuno affinity column,the chitinase in M.charantia leaves was isolated.The result of SDS PAGE showed that the Mr of the chitinase purified by immuno affinity chromatography was 35 kD,corresponding with that purified by chitin gel affinity chromatography,which indicated that the immunoaffinity for one plant chitinase could be used for purification of the same kind of chitinases in other plant species.Compared with chitin gel affinity,the immuo affinity chromatography was proved as a simple and fast way for chitinases purification when the antiserum was obtained,and the column could be used repeatedly. In vitro test,the chitinase purified through immuno affinity column was shown the ability in degrading hyphal wall preparation and inhibition to germ tube growth of cotton wilt pathogen, Fusarium oxysporum .
出处
《中国生物化学与分子生物学报》
CAS
CSCD
1999年第6期953-956,共4页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金
