| 甘蓝SRK激酶结构域编码区分子特性及其与THL1相互作用检测 |
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| 引用本文： | 高启国,宋明,牛义,杨昆,朱利泉,王小佳.甘蓝SRK激酶结构域编码区分子特性及其与THL1相互作用检测[J].农业生物技术学报,2008,16(5). |
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| 作者姓名： | 高启国 宋明 牛义 杨昆 朱利泉 王小佳 |
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| 作者单位： | 重庆北碚.西南大学园艺园林学院 重庆北碚.西南大学农学与生命科学学院 |
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| 摘 要： | 以甘蓝‘E1’为材料，采用PCR和RT-PCR技术，扩增SRK激酶结构域（记为SRKE1）编码区DNA和cDNA序列，得到片段长度分别为1711bp和1241bp，序列分析表明该序列由五个内含子和六个外显子组成，编码407个氨基酸；构建了SRKE1原核表达质粒pET43.1-SRKE1和真核表达质粒pPIC9K-SRKE1，分别在表达宿主菌大肠杆菌BL21和毕赤酵母GS115中进行表达。用我们表达的THL1与SRKE1孵育，结果显示SRKE1可与THL1结合并形成稳定的复合体。
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| 关 键 词： | 甘蓝 S-位点受体激酶 基因克隆 表达 |
| 收稿时间： | 2007-12-26 |
| 修稿时间： | 2008-5-30 |
Molecular Characterization of SRK Kinase-domain from Brassica oleracea L. and Assay of It’s Interaction with THL1 |
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| Abstract: | The DNA and cDNA fragments of SRK kinase domain(named as SRKE1) was amplified from stigma mRNA of Brassica oleracea L. by PCR and RT-PCR. Their lengths were 1241 bp and 1711 bp respectively. Sequence analysis indicated that the SRKE1 contained six exons and five introns，the SRKE1 encoded 407 amino acids. The coding sequence of SRKE1 were inserted into vector pET43.1 and pPIC9K to be the recombinant plasmid pET43.1-SRKE1 and pPIC9K-SRKE1. The recombinant protein were expressed in E. col(BL21) and Pichia pastoris GS115 repectively. When the SRKE1 was incubated with THL1 which we had expressed, SRKE1 could bind with THL1 and formed a stable complex. |
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