摘要
[目的]研究转基因棉田土壤微生物的功能多样性。[方法]采用直接法提取转基因棉根际土壤微生物总DNA,用限制性内切酶BamHI进行部分酶切,回收2.0～4.0 kb大小的片段插入到pUC19/BamHI脱磷载体上,转化到DH5α高效感受态细胞,构建宏基因组文库。随机挑取10个克隆序列测序,并通过NCBI预测可能存在的开放式阅读框和进行序列比对分析。[结果]该文库的外源插入片段平均长度为2.4 kb,90%以上克隆都含有插入片段,10个序列中存在着多种功能的开放式阅读框,包括一些水解酶、脱氢酶、脂解酶、修饰酶、抗生素及与电子传递链相关的酶类。[结论]该研究可为研究转基因棉大面积野外种植以后的生态安全性提供参考。
[Objective] The research aimed to study the functional diversity of microbes in the Bt cotton plot.[Method] The total DNA of microbes in rhizospheric soil of Bt cotton was extracted by the direct extraction method,and was partially digested by Bam HI.The fragments between 2.0 to 4.0 kb were recovered and inserted into pUC19/Bam HI vector,then transferred into DH5α,to construct the metagenomic library.DNA sequencing was conducted on 10 clones,then predicated maybe existing open reading frame(ORF) by NCBI and...
出处
《安徽农业科学》
CAS
北大核心
2010年第36期20559-20561,20599,共4页
Journal of Anhui Agricultural Sciences
基金
环保公益性行业科研专项(200709047)
江苏省自然科学基金项目(BK2006501)
关键词
转基因棉
宏基因组文库
未培养微生物
The Bt cotton
The metagenomic library
The functional uncultureable microorganism
