【摘要】 采用CTAB-SDS-冻融法和玻璃粉吸附法提取土壤宏基因组(metagenome),经琼脂糖凝胶电泳后用回收试剂盒对其进行纯化。以细菌16SrDNA通用引物从宏基因组中扩增出V8和V9两个高变区,回收纯化后与克隆载体pMD18-T连接,转化大肠杆菌DH5α,随机挑取一个阳性克隆菌进行序列测定,经BLAST分析表明该序列所代表的是一个不动杆菌属(Acinetobacter)细菌。本研究结果为分析土壤微生物种群结构和直接从土壤中分离功能基因奠定了基础。更多还原

【Abstract】 Two simple and rapid methods, which were CTAB-SDS-freezing-thawing and glass powder absorbing, were applied for metagenome preparation from soil samples. Ultraclean 15 DNA Purification Kit was used to purify the crude metagenome extract prepared by CTAB-SDS-freezing-thawing. PCR amplification for bacteria 16s rDNA by universal primers was achieved from the two metagenome preparations. The products were purified, then ligated into pMD18-T vector, and transformed into E.coli DH5α. One positive clone was sequenced. Blast analysis showed that the sequence was part of the 16S rDNA of Acinetobacter. This study provides foundation for microbial diversity analysis and direct cloning of functional genes from soil samples.更多还原