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以“华南8号”木薯(SC8)和“华南124号”木薯(SC124)的叶绿体作为研究材料,采用改进酚抽提法提取蛋白,通过单向SDS-PAGE电泳和双向SDS-PAGE电泳,比较不同木薯品种叶绿体的蛋白表达谱,并对表达的差异蛋白进行MALDI-TOF MS质谱鉴定,获得15个差异蛋白,其中有6个蛋白在SC124木薯叶绿体中表达较高,9个蛋白表达很低。对蛋白进行功能分析,发现差异蛋白主要参与蛋白翻译后修饰、周转、分子伴侣、碳水化合物运输等过程。通过RT-PCR验证了木薯核酮糖1,5-二磷酸羧化酶、ATP合酶β亚基的基因表达情况,结果表明,ATP合酶β亚基基因表达与蛋白质的表达比较一致,而核酮糖1,5-二磷酸羧化酶基因与蛋白质表达变化不一致。
The chloroplasts of SC8 and SC124 in South China 8 were used as materials to extract protein by improved phenol extraction method. SDS-PAGE electrophoresis and two-dimensional SDS- PAGE electrophoresis to compare the expression profiles of chloroplasts in different cassava varieties. The differential proteins were identified by MALDI-TOF MS, and 15 differential proteins were obtained. Among them, 6 proteins were highly expressed in chlorophyll of SC124 cassava and 9 proteins Low expression. Functional analysis of the protein found that the differential proteins are mainly involved in protein post-translational modification, turnover, chaperones, carbohydrate transport and other processes. The gene expression of ribulose 1,5-bisphosphate carboxylase and ATP synthase β subunit of cassava was verified by RT-PCR. The results showed that the expression of ATP synthase β subunit gene was consistent with the protein expression Ribulose 1,5-bisphosphate carboxylase gene and protein expression change inconsistent.