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当前位置: 首页 > 文章 > 甘蓝BoSU03蛋白基因的克隆及其与SRK相互作用分析 园艺学报 2015,42 (11) 2206-2214
Position: Home > Articles > Molecular Cloning of BoSU03 and Analysis of the Interaction Between SRK and BoSU03 in Brasscia oleracea Acta Horticulturae Sinica 2015,42 (11) 2206-2214

甘蓝BoSU03蛋白基因的克隆及其与SRK相互作用分析

作  者：
毕云龙;高启国;施松梅;刘晓欢;蒲全明;张莹;张林成;廉小平;柳菁
单  位：
西南大学农学与生物科技学院;西南大学园艺园林学院
关键词：
甘蓝;S位点受体激酶(SRK);BoSU03;植物泛素蛋白家族(PUB);下游信号蛋白;酵母双杂交系统
摘  要：
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。
译  名：
Molecular Cloning of BoSU03 and Analysis of the Interaction Between SRK and BoSU03 in Brasscia oleracea
作  者：
BI Yun-long;GAO Qi-guo;SHI Song-mei;LIU Xiao-huan;PU Quan-ming;ZHANG Ying;ZHANG Lin-cheng;LIAN Xiao-ping;LIU Jing;ZHU Li-quan;College of Agronomy and Biotechnology,Southwest University;College of Horticulture and Landscape Architecture,Southwest University,Key Laboratory in Olericulture of Chongqing;
关键词：
Brasscia oleracea;;S-locus receptor kinase(SRK);;BoSU03;;Plant U-box family(PUB);;downstream signaling protein;;yeast two-hybrid system
摘  要：
In this study,the coding sequences of ubiquitin protein BoSU03 identified by Brassica oleracea stigma protein mass spectrometry were amplified.Nucleotide sequence analysis showed that it was highly identical with nucleotide sequence of ubiquitin protein Bol003815 in Brasscia oleracea,BoSU03 contained ARM repeat domain and belonged to Plant U-box protein family.In order to analyze the interaction between BoSU03 and SRK,GAL4 two-hybrid system was used here,and the interaction between SRK and ARC1 was used as control.The results showed that the yeast cells transformed with BoSRK/BoSU03 could grow better and turn deeper blue on SD/-Ade/-His/-Leu/-Trp/x-α-gal/25 mmol 3-AT nutritional media.And the level of β-Galactosidase activity of BoSRK/BoSU03 was higher than that of BoSRK/BoARC1.These results will provide the basis for further analysis whether BoSU03 participates in SRK downstream SI signaling pathway.

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