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不结球白菜高亲和性硝酸盐转运蛋白NRT2.2的克隆和表达分析
引用本文:孙菲菲,;王镇,;李建刚,;王夏,;孙雪花,;侯喜林.不结球白菜高亲和性硝酸盐转运蛋白NRT2.2的克隆和表达分析[J].江西农业学报,2014(9):6-11.
作者姓名:孙菲菲  ;王镇  ;李建刚  ;王夏  ;孙雪花  ;侯喜林
作者单位:[1]江苏省南京市蔬菜科学研究所,江苏南京210042; [2]南京农业大学作物遗传与种质创新国家重点实验室,江苏南京210095; [3]中国科学院土壤环境与污染修复重点实验室、中国科学院南京土壤研究所,江苏南京210008
基金项目:国家自然科学基金项目(31201634、41201241); 江苏省自然科学基金项目(BK2012074); 博士后基金项目(2013M531412); 江苏省南京市科技计划项目(2013403S)
摘    要:以不结球白菜品种"苏州青"为材料,克隆得到了1个高亲和性硝酸盐转运蛋白基因——BcNRT2.2,该基因的全长为1593 bp,编码530个氨基酸。序列分析结果表明:BcNRT2.2基因含有AGWGNMG共识别基序,其氨基酸序列与拟南芥NRT2.2的同源性为86%;其二级结构主要由α-螺旋和自由卷曲构成;预测BcNRT2.2蛋白含有12个跨膜域。实时定量PCR分析结果表明:BcNRT2.2主要在根部表达,并受高浓度硝酸盐的诱导;BcNRT2.2在原始叶片中表达量较低,但受低温、高温和干旱胁迫的诱导后而高量表达。

关 键 词:不结球白菜  硝酸盐转运蛋白  克隆  表达  非生物胁迫

Cloning and Expression Analysis of Gene BcNRT2. 2 in Non- heading Chinese Cabbage
Institution:SUN Fei - fei, WANG Zhen, LI Jian - gang, WANG Xia , SUN Xue - hua , HOU Xi - lin. (1. Nanjing Vegetable Science Research Institute of Jiangsu Province, Nanjing 210042, China; 2. State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China; 3. Key Laboratory of Soil Environment and Pollution Remediation, Chinese Academy of Sciences; Nanjing Institute of Soil Science, Chinese Academy of Sciences, Nanjing 210008, China)
Abstract:In this study,non- heading Chinese cabbage( Brassica campestris ssp. chinensis Makino) cultivar"Suzhouqing"was used as experimental material,and a high- affinity nitrate transporter gene BcNRT2. 2 was successfully cloned. This gene had the total length of 1593 bp,and encoded 530 amino acids. The results of sequence analysis showed that the gene BcNRT2. 2 contained the identified sequence AGWGNMG of MFS super gene family,and the amino acid sequences of BcNRT2. 2 had 86% homology with those of NRT2. 2 in arabidopsis. The secondary structure of BcNRT2. 2 was mainly composed of alpha helices and random coil. It was predicted that the protein BcNRT2. 2 contained 12 transmembrane domains. Real- time quantitative PCR analysis indicated that the expression of BcNRT2. 2 was found mainly in the roots,and was induced by high concentration of nitrate; the expression quantity of BcNRT2. 2 in original leaf was relatively low,but abiotic stress( low temperature,high temperature,and drought)could rapidly induce its high- quantity expression.
Keywords:Non-heading Chinese cabbage  Nitrate transporter protein  Cloning  Expression  Abiotic stress
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