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大豆GmDRR与互作基因GmDIP1的亚细胞定位及原核表达
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作者 杨蕊 禹国龙 +4 位作者 沈岩茹 刘春燕 傅永福 辛大伟 陈庆山 《大豆科学》 CAS CSCD 北大核心 2016年第4期557-561,共5页
本实验室通过疫霉菌诱导大豆绥农10号后,获得差异表达蛋白点,通过序列拼接比对和分析,获得一个新的大豆GmDRR基因,并通过酵母双杂筛选文库的方式获得其互作基因GmDIP1。为进一步明确两个基因关系,对两个基因分别进行亚细胞定位和原核表... 本实验室通过疫霉菌诱导大豆绥农10号后,获得差异表达蛋白点,通过序列拼接比对和分析,获得一个新的大豆GmDRR基因,并通过酵母双杂筛选文库的方式获得其互作基因GmDIP1。为进一步明确两个基因关系,对两个基因分别进行亚细胞定位和原核表达以及重组蛋白的纯化。GmDRR蛋白为外分泌蛋白,定位于细胞膜与细胞质中,GmDIP1基因定位于线粒体膜上,二者都成功进行了原核表达,从而获得了有表达功能和生物学活性的蛋白。 展开更多
关键词 大豆 gmdrr GmDIP1 亚细胞定位 原核表达
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GmDRR1,a dirigent protein resistant to Phytophthora sojae in Glycine max (L.) Merr.
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作者 CHEN Qing-shan YU Guo-long +15 位作者 ZOU Jia-nan WANG Jing QIU Hong-mei ZHU Rong-sheng CHANG Hui-lin JIANG Hong-wei HU Zhen-bang LI Chang-yu ZHANG Yan-jiao WANG Jin-hui WANG Xue-ding GAO Shan LIU Chun-yan QI Zhao-ming FU Yong-fu XIN Da-wei 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2018年第6期1289-1298,共10页
Soil-borne pathogen Phytophthora sojae is an oomycete that causes devastating damage to soybean yield. To mine original resistant genes in soybean is an effective and environmentally-friend approach controlling the di... Soil-borne pathogen Phytophthora sojae is an oomycete that causes devastating damage to soybean yield. To mine original resistant genes in soybean is an effective and environmentally-friend approach controlling the disease. In this study, soybean proteins were extracted from the first trifoliolates infected by predominant P. sojae race 1 and analyzed by twodimensional gel electrophoresis. Nineteen differently-expressed protein spots were detected, and 10 of them were further applied for Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry Assay. One protein containing a dirigent (DIR) domain was identified and belonged to the DIR-b/d family. Therefore, it was named as GmDRR1 (Glycine max Disease Resistance Response 1). Then, GmDRR1 gene was pathologically confirmed to be involved in the resistant to P. sojae in soybean. GmDRR1-GFP (green fluorescent protein) fusion proteins localized in the cell membrane. qRTPCR results showed GmDRR1 gene expressed differently in P. sojae resistant- and susceptible-soybean cultivars. By the promoter analysis, we found a haplotype H8 was existing in most resistant soybean varieties, while a haplotype H77 was existing in most susceptible soybean varieties. The H77 haplotype had seven SNPs (C to A, G to C, C to A, T to A, T to C, T to C, and T to A) and two single nucleotide insertions. The results supported that the expression difference of GmDRR1 genes between P. sojae resistant- and susceptible-soybean cultivars might depend on the GmDRR1 promoter SNPs. The results suggested that GmDRR1 was a dirigent protein involved in soybean resistant to P. sojae and paved a novel way for investigation of the molecular regulatory mechanism of the defense response to P. sojae in soybean. 展开更多
关键词 dirigent domain gmdrr1 SOYBEAN Phytophthora sojae OOMYCETE PROTEOMICS
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