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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 order to determine whether the molecular weight of polygalacturonase is the same between the Japanese pear and the European pear, Polygalacturonase gene amplification, sequencing and comparison , The similarity is 96%. To further determine whether the two polygalacturonase proteins in the subsequent processing are different, the further use of extension obtained polygalacturonase partial cDNA sequence, and the use of plasmid pET-24a (+) pairs The obtained cDNA sequence expression, Ni-NTA column purification of recombinant protein, mouse immunodeficiency virus and other methods to obtain polygalacturonase pear polygalacturonase antibody by Western blotting method of pear pear and pear European black Aspergillus niger polygalacturonase molecular weight ratio, the results show that the two pear scab polygalacturonase molecular weight there is no difference.