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扩增了从甘蓝柱头蛋白质谱鉴定中获得的泛素蛋白BoSU03基因的编码序列。序列分析表明BoSU03与甘蓝泛素蛋白Bo1003815的核苷酸序列相似性最高,达98%,含有臂重复结构域,属于植物泛素蛋白家族。为探究BoSU03是否为自交不亲和S位点受体激酶(SRK)下游的互作蛋白,以SRK与ARC1相互作用为对照,利用GAL酵母双杂交系统对SRK与BoSU03进行了相互作用验证,结果表明,BoSRK/BoSU03的转化酵母在四缺平板SD/-Ade/-His/-Leu/-Trp/x-α-gal/25 mmol 3-AT上长势较好,显色较深,进一步检测发现BoSRK/BoSU03转化酵母的β-半乳糖苷酶活性值大于BoSRK/BoARCl转化酵母的活性值,这为进一步分析鉴定BoSU03是否参与SRK下游信号传导过程提供了依据。
The coding sequence of the ubiquitin protein BoSU03 gene obtained from identification of cabbage stigma protein was amplified. Sequence analysis showed that the nucleotide sequence similarity of BoSU03 and Bo1003815 was 98%, which contained arm repeat domain and belonged to plant ubiquitin protein family. To investigate whether BoSU03 is an interaction protein downstream of self-incompatibility S-site receptor kinase (SRK), SRK interacts with BoSU03 by using the GAL yeast two-hybrid system with the interaction between SRK and ARC1 as the control, The results showed that the transformed yeast of BoSRK / BoSU03 grew well on the four-deficient plate SD / -Ade / -His / -Leu / -Trp / x-α-gal / 25 mmol 3-AT, It was found that the β-galactosidase activity of BoSRK / BoSU03-transformed yeast was higher than that of BoSRK / BoARCl-transformed yeast, which provided the basis for further analysis to identify whether BoSU03 participates in the SRK downstream signaling.