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]
抗2,4-D巨大芽孢杆菌基因组文库的构建
- Title:
- Genomic Library Construction of Bacillus Megaterium Resistant to 2,4-D
- 文章编号:
- 1000-9841.2014.06.0826
- Keywords:
- 2; 4-D resistant; Bacillus megaterium; Genomic library; Transgenic soybean
- 分类号:
- Q933
- 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:
[1]李文霞,李柏云,薛红,等.黑龙江省不同生态区大豆品种育种性状的主要成分分析[J].大豆科学,2013,32(6):731-734.(Li W X,Li B Y,Xue H,et al.Principal components analysis of breeding traits in various ecological regions in Heilongjiang province[J].Soybean Science,2013,32(6):73-76.)
备注/Memo
基金项目:转基因生物新品种培育重大专项(2008ZX08004-001)。