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本研究利用表达油菜波里马胞质雄性不育相关线粒体基因 orf224 3′端(B)片段的大肠杆菌,通过分级分离纯化得到了 GST-B 蛋白,并和亲和层析纯化的 GST 蛋白一起分别制备了抗血清。利用纯化的抗血清对在 pBV220和 pGEX-5X-1系统表达的 orf224各基因片段产物进行了分析,结果表明纯化的 GST-B 抗体具有 ORF224蛋白专一性,同时也证实了 orf224全基因及其3’端片段的表达。研究结果为进一步分析波里马不育胞质线粒体中 orf224基因的表达奠定了基础。
In this study, GST-B protein was isolated and purified by fractionating and purifying Escherichia coli expressing the 3 ’end (B) fragment of orf224 related to mitochondrial DNA gene of B. napus. Antisera were prepared. Purified antiserum was used to analyze the orf224 gene fragments expressed in pBV220 and pGEX-5X-1 system. The results showed that the purified GST-B antibody has ORF224 protein specificity, and also confirmed that orf224 whole gene and its 3 ’end of the expression of the fragment. The results laid the foundation for the further analysis of orf224 gene expression in Polymorphic sterile rice.