[1]张秀春　郭丽琼**,吴坤鑫　林俊扬　林俊芳**.双 T - DNA 表达载体转化大豆的研究*[J].大豆科学,2005,24(04):291-295.[doi:10.11861/j.issn.1000-9841.2005.04.0291]
　Zhang Xiuchun Gou Liqiong *Wu Kunxin Lin Junyang Lin Junfang.AGROBACTERIUM - MEDIATED TRANSFORATION OF SOYBEAN WITH THE EXPRESSIONVECTOR CARRYING TWO SEPARATE T- DNAs[J].Soybean Science,2005,24(04):291-295.[doi:10.11861/j.issn.1000-9841.2005.04.0291]
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双 T - DNA 表达载体转化大豆的研究*

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第24卷 期数: 2005年04期 页码: 291-295 栏目: 出版日期: 2005-11-25
Title:
AGROBACTERIUM - MEDIATED TRANSFORATION OF SOYBEAN WITH THE EXPRESSIONVECTOR CARRYING TWO SEPARATE T- DNAs
作者:
张秀春1　郭丽琼**1;  2　吴坤鑫1　林俊扬3　林俊芳**1;  2
(1. 中国热带农业科学院热带生物技术研究所,热带生物技术国家重点实验室 ,海口 571101;2. 华南农业大学食品学院生物工程系 ,广州 510640;3. 福建省莆田市农业科学研究所, 莆田 351100)
Author(s):
Zhang Xiuchun 1 　Gou Liqiong 1; 2 *Wu Kunxin 1 　Lin Junyang 3Lin Junfang 1; 2
(1. State Key Laboratory of Tropical Crop Biotechnology , Chinese Academy of TropicalAgricultural Sciences. Haikou 571101;2.Department of Bioengineering , College of FoodSciences,South China Agricultural University . Guangzhou 510640;3. Putian City’sInstitute of Agricultural Sciences. Putian,351100)
关键词:
共转化表达载体 ; Δ6- 脂肪酸脱氢酶基因 ; 转基因大豆
Keywords:
Co - transformation expression vector; Δ6- desaturase gene ; Transgenic soybean
DOI:
10.11861/j.issn.1000-9841.2005.04.0291
文献标志码:
A
摘要:
利用含有筛选标记基因和目的基因 Δ6- 脂肪酸脱氢酶基因的双 T - DNA 共转化表达载体pLIN61,经农杆菌 EHA101 介导,采用子叶节转化法转化大豆, 使用除草剂 Glufosinate 作为筛选剂,获得了一批携带有玻璃苣 Δ6- 脂肪酸脱氢酶基因的转基因大豆, 转化效率为 2. 0%, 共转化频率及实际转化率分别为 56. 25%、1. 12%。PCR 检测和 Southern 杂交结果证明 ,玻璃苣 D6D 已经整合到大豆的基因组上。RT - PCR检测结果显示, 玻璃苣 Δ6- 脂肪酸脱氢酶基因在转基因大豆的转录水平上得到了表达 。
Abstract:
The co - transformation expression vector , pLIN61 carrying two separate T - DNAs with select-able marker gene and Δ6- desaturase gene was used to transformed soybean cotyledonary nodes mediatedby Agrobacterium tumef aciens strain EHA101. A batch of transgenic soybeans harboring Δ6- desaturasegene were obtained through Glufosinate screening. Research results demonstrate that the transformationefficiency , co - transformation frequency and real transformation frequency wrer 2. 0%, 56. 25% and1. 12 %respectively. PCR detection and Southern analysis confirm that borage Δ6- desaturase gene Gas in-tegrated into soybean genome. RT - PCR detection showed that borage Δ6- desaturase gene had expressedat transcriptional level in the transgenic soybeans.

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备注/Memo

基金项目:国家自然科学基金项目(30371000), 广东省自然科学基金项目(032239), 广东省科技攻关项目(2005B210201009)和华南农业大学校长基金项目(5100- K03003)

作者简介:张秀春(1972 -), 女, 讲师, 在读博士生, 主要从事植物生物技术工作。**郭丽琼为并列
第一作者, 林俊芳为通讯作者。E. manil:linjf @scau. edu. cn

更新日期/Last Update: 2017-01-13