[1]徐思靓,韩潮,金龙国,等.抗2,4-D巨大芽孢杆菌基因组文库的构建[J].大豆科学,2014,33(06):826-829.[doi:10.11861/j.issn.1000-9841.2014.06.0826]
　XU Si-liang,HAN Chao,JIN Long-guo,et al.Genomic Library Construction of Bacillus Megaterium Resistant to 2,4-D[J].Soybean Science,2014,33(06):826-829.[doi:10.11861/j.issn.1000-9841.2014.06.0826]
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抗2,4-D巨大芽孢杆菌基因组文库的构建

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第33卷 期数: 2014年06期 页码: 826-829 栏目: 出版日期: 2014-12-25
Title:
Genomic Library Construction of Bacillus Megaterium Resistant to 2,4-D
文章编号:
1000-9841.2014.06.0826
作者:
徐思靓1; 2; 韩潮1; 2; 金龙国2; 邱丽娟2; 陶波1
1.东北农业大学 农学院，黑龙江 哈尔滨 150030；
2.中国农业科学院 作物科学研究所/国家农作物基因资源与遗传改良重大科学工程/农业部作物种质资源与生物技术重点开放实验室，北京 100081
Author(s):
XU Si-liang1; 2; HAN Chao1; 2; JIN Long-guo2; QIU Li-juan2; TAO Bo1
1.College of Agriculture,Northeast Agricultural University,Harbin 150030，China;
2.The National Key Facility for Crop Gene Resources and Genetic Improvement(NFCRI)/Key Laboratory of Germplasm & Biotechnology(MOA),Institute of Crop Sciences,Chinese Academy of Agricultural Sciences,Beijing 100081,China
关键词:
抗2; 4-D; 巨大芽孢杆菌; 基因组文库; 转基因大豆
Keywords:
2; 4-D resistant; Bacillus megaterium; Genomic library; Transgenic soybean
分类号:
Q933
DOI:
10.11861/j.issn.1000-9841.2014.06.0826
文献标志码:
A
摘要:
以抗2,4-D的巨大芽孢杆菌Bacillus megaterium为材料，酶切基因组总DNA和载体pACYC184；回收1~4 kb范围的外源基因组DNA片段，与载体连接后，转入BL21感受态细胞中,进行蓝白斑与氯霉素抗性筛选,采用菌落PCR方法检测连接效率,并对该文库进行质量鉴定。成功构建了巨大芽孢杆菌基因组文库，得到8 900个阳性重组子，平均插入片段长度为2.5 kb，文库约覆盖了B.megaterium菌株基因组的556倍。该文库的建立为选育抗2，4-D转基因大豆的新品种奠定了基础。
Abstract:
Bacillus megateriumwhich was efficient resistant to 2,4-D,recycled the 1~4 kb DNA fragments of digested B.megaterium genomic DNA,and with digested vector pACYC184 overnight connection,transferred it to BL21 competent cells,after the blue-white screening and chloramphenicol resistance screening by colony PCR to detect the connection efficiency and quality of the library.Successfully constructed the B.megaterium genomic library and obtain 8 900 positive clones with an average insert fragment length of 2.5 kb.And this library covers approximately 556 times B.megaterium genome.This library establish the theoretical basis for 2,4-D resistant transgenic soybean.

参考文献/References:

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备注/Memo

基金项目：转基因生物新品种培育重大专项(2008ZX08004-001)。

第一作者简介：徐思靓（1989-），女，硕士，主要从事除草剂抗性基因研究。E-mail：15045116795@139.com。

更新日期/Last Update: 2014-12-29