重金属污染土壤中泛基因组的提取及其细菌种类的免培养法分析  

Isolation for the metagenome from the soil sample polluted by heavy metal and bacterial analysis by culture-free method

作　　者：林毅[1] 洪雪梅[2] 蔡丽希[1] 方光伟[1] 彭锟[1] 

机构地区：[1]华侨大学生物工程与技术系,福建泉州362021 [2]华侨大学信息科学与工程学院,福建泉州362021

出　　处：《漳州师范学院学报（自然科学版）》2005年第1期56-59,共4页Journal of ZhangZhou Teachers College（Natural Science)

基　　金：福建省青年科技人才创新项目(2003J024)

摘　　要：采用玻璃粉吸附法提取重金属污染土壤的泛基因组,利用细菌16SrDNA保守区段B2/B3为引物,经PCR扩增获得包含V8和V9两个高变区的大小为1050bp的产物.将回收纯化的PCR产物与克隆载体pMD18-T连接,转化大肠杆菌DH5α感受态细胞,经抗生素抗性和蓝白斑筛选,并提取质粒作酶切分析,随机挑取3个阳性克隆菌进行序列测定,结果表明3个序列所代表的均是芽孢杆菌属(Bacillus)细菌,而且均是与Bacillusflexus亲缘关系非常接近的细菌.Metagenome was extracted from the soil sample polluted by heavy metal using SiO2 absorbing. The 1050 bp PCR product was amplified by conserved primers for bacterial 16S rDNA using the metagenome as template. After purified, the product was ligated into pMD18-T vector, then transformed into E.coli DH5α. The sequences of the three positive clones all showed close relationships to Bacillus Flexus.

关 键 词：细菌种类 基因组 抗生素 DH 白斑 克隆载体 质粒 土壤 区段 亲缘关系 

分 类 号：S826[农业科学—畜牧学]