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甘蓝SRK的S域及SCR酵母表达载体的构建及其相互作用区段的研究 被引量：1

Studies on the Construction of S Domain of SRK and SCR Gene's Yeast Expression Vectors and Their Interaction in Yeast Two-hybrid System
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摘要 为了深入研究S位点受体激酶（SRK）和S位点富含半胱氨酸蛋白（SCR）甘蓝自交不亲和（self-incompatibility,SI）信号传导的雌雄决定因子的相互作用机制,以自交不亲和性高的结球甘蓝为材料,采用酵母双杂交体系,构建pGADT7-eSRKs和pGBKT7-SCRs重组载体,并分别转化到Y187和Y2HGold酵母中,通过SD/-Ade-His-Trp-Leu平板上菌落的形成,PCR以及x-α-gal显色反应鉴定转化到酵母中的两个重组质粒相互作用。结果表明,SRK S域的SRK1及SRK4分别与SCR2相互作用,且初步显示其相互作用区域为SRK第1个外显子的第16~421bp的片段和SCR第1876~2068bp的片段。这既为SRK-SCR相互作用提供了具体证据,也为甘蓝自交不亲和性分子机理的深入研究提供了新内容。 For further study on the interaction of SRK-SCR and between these two determinant factors,Brassica oleracea with high self-incompatibility was used as research materials to construct a group of pGADT7-eSRKs and pGBKT7-SCRs recombinant vectors that were subsequently transformed accordingly into Y187 and Y2HGold yeast.The interaction between these two recombinant plasmids transformed into the yeasts were tested by SD/-Ade-His-Trp-Leu fungal colony formation of plates,PCR and x-α-gal color reactions.The results showed that there existed two kinds of interactions among all interaction assays,which preliminary indicated that their interaction region span 16-421 bp fragment of the first exon of SRK and 1 876-2 068 bp fragment of SCR.These results reconfirm the SRK-SCR interaction,adding some insights into the mechanism of selt incompatibility in Brassica.
作者 彭一波 朱利泉 杨红 薛丽琰 罗兵 吴志刚 黄丹 杨昆 高启国 李成琼 任雪松 王小佳
机构地区 西南大学农学与生物科技学院 重庆市蔬菜学重点实验室
出处 《园艺学报》 CAS CSCD 北大核心 2011年第8期1479-1488,共10页 Acta Horticulturae Sinica
基金 国家自然科学基金项目(30971849) 高校基本科研业务专项基金项目(XDJKC109)
关键词 甘蓝 SRK eSRKs基因 SCRs基因 pGADT7载体 pGBKT7载体 酵母双杂交体系 Brassica oleracea SRK eSRKs gene SCRs gene pGADT7 vector pGBKT7 vector yeast two-hybrid system
分类号 S635 [农业科学—蔬菜学]
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参考文献13

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二级参考文献15

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共引文献27

同被引文献17

  • 1郑斌,詹希美.信号肽序列及其在蛋白质表达中的应用[J].生物技术通讯,2005,16(3):296-298. 被引量：45
  • 2芦岩,李玉花.芸薹属中自交不亲和反应的信号转导[J].植物生理学通讯,2005,41(4):547-552. 被引量：5
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  • 8Takayama S, Shimosato H, Shiba H, Funato M, Che F S, Wata- nabe M, Iwano M, Isogai A. Direct ligand-receptor complex in- teraction controls Brassica self-incompatibility. Nature, 2001, 413:534-538.
  • 9Mishima M, Takayama S, Sasaki K I, Jee J G, Kojima C, Isogai A, Shirakawa M. Structure of the male determinant factor for Bras- sica self-incompatibility. J Biol Chem, 2003, 278:36389-36395.
  • 10YangH(杨红),ZhuL-Q(朱利泉),ZhangH-C(张贺翠),YangY-J(杨永军),XueL-Y(薛丽琰),YangK(杨昆),YuH(余浩),PengY-B(彭一波),LuoB(罗兵),WuZ-G(吴志刚),HuangD(黄丹),GaoQ-G(高启国),WangX-J(王小佳).Study on the interac-tions between the truncated fragments of SCR and eSRK from Brassica oleracea L. by a yeast two-hybrid system.中国农业科学,2011,44(9):1953-1962(in Chinese with Eng-1ish abstract).

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园艺学报
2011年 第8期

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