日本梨黑星病菌与欧洲梨黑星病菌多聚半乳糖醛酸酶研究

Studies on the polygalacturonases produced by pear scab pathogens,Venturia nashicola and Venturia pirina

为判断日本梨黑星病菌与欧洲梨黑星病菌多聚半乳糖醛酸酶蛋白分子量是否相同,本研究对2种梨黑星病菌多聚半乳糖醛酸酶进行基因扩增、测序与比对,相似度为96%。为进一步判断两者多聚半乳糖醛酸酶的蛋白在后续加工过程中是否有所差异,进一步利用延伸法获得多聚半乳糖醛酸酶部分cDNA序列,并利用质粒pET-24a(+)对获得的cDNA序列表达、Ni-NTA柱纯化重组蛋白、用蛋白免疫小鼠等方法获得梨黑星病菌多聚半乳糖醛酸酶抗体,通过Western blotting方法进行了日本梨黑星病菌与欧洲梨黑星病菌多聚半乳糖醛酸酶分子量比对,结果证明2种梨黑星病菌的多聚半乳糖醛酸酶分子量没有区别。

In ordor to determine whether the molecular weight of Venturia nashicola and Venturia pirina were same.Polygalacturonases gene of V.nashicola and V.pirina were cloned,sequenced and matched.The similarity was 96%.Partial cDNA sequence of polygalacturonase obtained using extension method in this study.The polygalacturonases cDNA sequence of V.nashicola obtained was expressed by plasmid pET-24a（＋）.Recombinant protein was purified by Ni-Chelated column.Antibody was got by immune mice.Molecular weight of polygalacturonases produced by V.nashicola and V.pirina was detected by western blotting,the result is that the molecular weight of polygalacturonases produced by V.nashicola and V.pirina was no difference.