[1]高明潇,郭娜,薛晨晨,等.大豆△1-吡咯啉-5-羧酸合成酶基因GmP5CS1和GmP5CS2的克隆与过表达载体构建[J].大豆科学,2016,35(01):25-30,38.[doi:10.11861/j.issn.1000-9841.2016.01.0025]
　GAO Ming-xiao,GUO Na,XUE Chen-chen,et al.Cloning of the Soybean △1-Pyrroline-5-Carboxylate Genes GmP5CS1 and GmP5CS2 and Construction of Its Overexpression Vectors[J].Soybean Science,2016,35(01):25-30,38.[doi:10.11861/j.issn.1000-9841.2016.01.0025]
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大豆△1-吡咯啉-5-羧酸合成酶基因GmP5CS1和GmP5CS2的克隆与过表达载体构建

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第35卷 期数: 2016年01期 页码: 25-30,38 栏目: 出版日期: 2016-01-20
Title:
Cloning of the Soybean △1-Pyrroline-5-Carboxylate Genes GmP5CS1 and GmP5CS2 and Construction of Its Overexpression Vectors
作者:
高明潇; 郭娜; 薛晨晨; 王海棠; 赵晋铭; 邢邯
南京农业大学 大豆研究所/国家大豆改良中心/农业部大豆生物学与遗传育种重点实验室/作物遗传与种质创新国家重点实验室，江苏 南京 210095
Author(s):
GAO Ming-xiao;  GUO Na;  XUE Chen-chen;  WANG Hai-tang;  ZHAO Jin-ming;  XING Han
Soybean Research Institute of Nanjing Agricultural University/National Center for Soybean Improvement/Key Laboratory of Biology and Genetics and Breeding for Soybean, Ministry of Agriculture/State Key Laboratory for Crop Genetics and Germplasm Enhancement, Nanjing 210095, China
关键词:
大豆; GmP5CS基因; Gateway技术
Keywords:
Glycine max L Merr;  GmP5CS gene;  Gateway technology
DOI:
10.11861/j.issn.1000-9841.2016.01.0025
文献标志码:
A
摘要:
利用同源克隆方法从强耐旱栽培大豆（Glycine max）品种齐黄22中克隆得到2个编码大豆△1-吡咯啉-5-羧酸合成酶（P5CS）的基因，分别命名为GmP5CS1和GmP5CS2。氨基酸序列分析发现，GmP5CS1包含1个长度为2 163 bp的ORF，编码720个氨基酸，等电点pI为6,70，分子量大小为78,38 kDa；GmP5CS2包含1个长度为2 271 bp的ORF，编码756个氨基酸，等电点pI为6.10，分子量大小为82.18 kDa。与NCBI公布的部分物种蛋白质序列比对发现，GmP5CS1与紫花苜蓿（Medicago sativa）的亲缘关系最高，GmP5CS2与豇豆（Vigna unguiculata）的亲缘关系最高。组织表达分析表明，GmP5CS1与GmP5CS2基因在大豆的各个组织中均有表达，其中叶和根中的表达量相对较高，茎中表达量次之，花和籽粒中的表达量相对较低。利用Gateway技术得到植物过表达载体pEarleyGate103-GmP5CS，再转入根癌农杆菌EHA105中，为GmP5CS基因的转化及功能分析奠定了基础。
Abstract:
In this study, we cloned two genes encoding soybean △ 1-?pyrroline-5- carboxylic acid synthase(P5CS) by homology cloning method of cultivation from the strong drought tolerant soybean (Glycine max) variety Qihuang 22, named GmP5CS1 and GmP5CS2. The amino acid sequence analysis found that GmP5CS1 contained a length of 2 163 bp ORF, encoding 720 amino acids, isoelectric point (pI) was 6.70, protein was weak acid, molecular weight was 78.38 kDa; ?GmP5CS2 contained a length of 2 271 bp ORF, encoding 756 amino acids, isoelectric point(pI) was 6.10, acidic protein, molecular weight was 82.18 kDa. Phylogenetic analysis showed that GmP5CS1 affinity with Medicago sativa highest, GmP5CS2 affinity with Vigna unguiculata highest The organization expression analysis found that GmP5CS1 and GmP5CS2 genes expressed in all soybean organizations.Gene relative expression in leaf and root is higher, relative expression in the stem is middle, the relative expression of flowers and seed is relatively lower A plant over-expression vector of pEarleyGate103-GmP5CS was constructed by Gateway technology, and was transferred into Agrobacterium EHA105, which would lay a foundation of its conversion into the plant and provide a foundation for further studying the function of GmP5CS gene in soybean

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备注/Memo

基金项目：国家转基因生物新品种培育科技重大专项(2014ZX08004)；国家自然科学基金(31301340，31301343)；国家现代农业产业技术体系(CARS-004-PS10)；江苏省农业科技支撑计划(BE2013350)；长江学者和创新团队发展计划(PCSIRT13073)；江苏省现代作物生产协同创新中心（JCIC-MCP）。

第一作者简介：高明潇（1989-），男，硕士，主要从事大豆分子遗传育种研究。E-mail：gaomingxiao1989@163.com。
通讯作者：邢邯（1963-），男，教授，博导，主要从事大豆育种工作。E-mail：hanx@njau.edu.cn。

更新日期/Last Update: 2016-01-28