WANG Bin,XU Zhihua,HOU Jinfeng,et al.Cloning of Soybean Cytoplasmic Invertase Gene GmCInv and Construction of Its Plant Expression Vector[J].Soybean Science,2014,33(02):168-172.[doi:10.11861/j.issn.1000-9841.2014.02.0168]
大豆细胞质转化酶基因GmCInv的克隆与植物表达载体构建
- Title:
- Cloning of Soybean Cytoplasmic Invertase Gene GmCInv and Construction of Its Plant Expression Vector
- 文章编号:
- 1000-9841（2014）02-0168-05
- 分类号:
- S565.1
- 文献标志码:
- A
- 摘要:
- 利用同源克隆方法从南农菜豆6号中克隆得到一个编码大豆细胞质转化酶(cytoplasmic invertase，CInv)的基因，命名为GmCInv。序列分析发现该基因含有一个长1 956 bp的开放阅读框，编码652个氨基酸。预测其蛋白质分子量为73.88 kDa，等电点为6.12。与NCBI公布的部分蛋白质序列比对发现，GmCInv蛋白序列与毛果杨(Populus trichocarpa)的相似性最高，达88%。组织表达分析表明，GmCInv在大豆苗期的叶、开花期的花中表达量最高，在根、茎中表达量较低。利用Gateway技术得到植物过表达载体pEarleyGate103GmCInv，再转入根癌农杆菌EHA105中，为GmCInv基因的转化及功能分析奠定了基础。
- Abstract:
- nvertase(INV),also known as sucrase is a hydrolytic enzyme that catalyzed irreversibly hydrolysis of sucrose to fructose and glucose,providing the growth and development of the plant with carbon and energy.Cytoplasmic invertase gene was isolated from vegetable soybean Nannong 6 by homology cloning,designed as GmCInv.GmCInv contained a 1 956 bp open reading frame,encoding a 652 amino acids protein.The protein is estimated to have a molecular weight of 73.88 kDa and isoelectric point of 6.12.Comparing with the amino acid sequence of CInv from other species on NCBI,GmCInv has a highest similarily of 88% with the CInv from Populus trichocarpa.The mRNA abundance of GmCInv in leaves of seedling stage and in flowers of florescence stage were higher than in stems and roots of the both stages.A plant over expression vector of pEarleyGate103GmCInv 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 GmCInv genein soybean.
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备注/Memo
基金项目：国家转基因重大专项(2012ZX08004)；国家自然科学基金(0106J1050)；现代农业产业技术体系建设专项(CARS 04 PS10)。
第一作者简介：王彬(1987-)，男，在读硕士，主要从事大豆分子遗传育种研究。E-mail：basibin2010@163.com。
通讯作者：邢邯(1963-)，男，教授，博士生导师，主要从事大豆育种研究。E-mail：hanx@njau.edu.cn。