[1]曲姗姗,刘丽君,寇坤,等.农杆菌介导法将Bt(cryⅠA)基因导入大豆的研究[J].大豆科学,2009,28(02):186-190，194.[doi:10.11861/j.issn.1000-9841.2009.02.0186]
　QU Shan-shan,LIU Li-jun,KOU Kun,et al.Transformation of -Bt(cryⅠA)-Gene into Soybean via Agrobacterium-mediated Method[J].Soybean Science,2009,28(02):186-190，194.[doi:10.11861/j.issn.1000-9841.2009.02.0186]
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农杆菌介导法将Bt(cryⅠA)基因导入大豆的研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第28卷 期数: 2009年02期 页码: 186-190，194 栏目: 出版日期: 2009-04-25
Title:
Transformation of -Bt(cryⅠA)-Gene into Soybean via Agrobacterium-mediated Method
文章编号:
1000-9841(2009)02-0186-05
作者:
曲姗姗1; 2; 刘丽君1; 寇坤1; 2; 唐晓飞1; 杨喆1
1黑龙江省农业科学院大豆研究所，黑龙江 哈尔滨 150086；
2东北农业大学研究生院，黑龙江 哈尔滨 150030
Author(s):
QU Shan-shan1; 2; LIU Li-jun1; KOU Kun1; 2; TANG Xiao-fei1; YANG Zhe1
1Soybean Research Institute，Heilongjiang Academy of Agricultural Sciences，Harbin 150086，Heilongjiang；
2Graduate School of Northeast Agricultural University，Harbin 150030，Heilongjiang，China
关键词:
大豆; Bt基因; 遗传转化
Keywords:
Soybean; Bt gene; Genetic transformation
分类号:
S565.1
DOI:
10.11861/j.issn.1000-9841.2009.02.0186
文献标志码:
A
摘要:
构建含有Bt（cryⅠA）基因的植物表达载体pCAMBIA3300-Bt，以大豆子叶节为受体，通过农杆菌介导法将Bt基因导入大豆品种黑农37中，获得转基因植株。并进行大豆的再生和遗传转化系统优化的研究，以获得较高的转化率。结果表明：在6-BA浓度为1.7 mgL-1时，丛生芽分化率最高。确定该品种大豆在丛生芽分化阶段的草铵膦筛选浓度为3.5 mgL-1。获得转化质粒pCAMBIA3300-Bt的转基因植株，其中T1代PCR阳性植株19株。采用real-time PCR的方法对T1代抗性植株进行Bt基因的转录水平的分析，初步证明Bt基因已整合到受体大豆的基因组内。
Abstract:
It is very important for diseases and insect pests controlling to transfer some resistant genes to obtain transgenic resistant varieties.In this study，plant expression vector pCAMBIA3300-Bt containing -Bt（cryⅠA）- gene was constructed，and transgenic plants were obtained by Agrobacterium-mediated transformation of soybean(Heinong 37) cotyledonary nodes.We regenerated plants and optimized the conditions of transformation to increase the efficiency of transformation.It had the optimal rate of shooting when the concentration of 6-BA was 1.7 mgL-1.The optimal selection concentration of Glufosinate-ammonium for shoot initiation was 3.5 mgL-1Transformed plants containing pCAMBIA3300-Bt was obtained and 19 positive plants from T1generation were tested by PCR.Real-time PCR was used to investigate the transcription level of Bt gene in T1transgenic plants，it confirmed the integration of Bt genes into the genome of soybean.

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备注/Memo

基金项目：国家高技术研究发展计划（863计划）资助项目（2006AA1021F9）；国家转基因重大专项资助项目（2008ZX08004-002）。

作者简介：曲姗姗(1983-)，女，硕士研究生，研究方向为植物基因工程与分子生物学。E-mail：qss3344@126.com。

更新日期/Last Update: 2014-10-03