%0 Journal Article
%T EXTRACTION OF DNA FROM ENVIRONMENTAL SAMPLES AND CONSTRUCTION OF MIXED GENOMIC DNA LIBRARIES<br>环境样品中DNA的分离纯化和文库构建
%A X Wang
%A Y Tang
%A J Wang
%A Y Huang
%A R Chen
%A L Huang
%A <br>王啸波
%A 唐玉秋
%A 王金华
%A 黄仪秀
%A 陈润生
%A 黄力
%J 微生物学报
%D 2001
%I 
%X A method has been developed for extracting and purifying genomic DNA from environmental samples. In this method, an environmental sample is treated first by grinding and freezing/thawing and subsequently by SDS/proteinase K-based DNA extraction. The yields of purified DNA from three samples used in this study ranged from 2 to 16 micrograms per gram of dry sample. Mixed genomic DNA libraries for two of the environmental samples were constructed by inserting restriction fragments (3-8 kb) of the purified DNAs into plasmid pUC18 and transforming E. coli DH5 alpha with the resultant plasmids. Approximately 10(3) to 10(4) insert-containing clones were obtained from 1 g of each sample. Clone libraries were analyzed by DNA sequencing and gene annotation. Among 20 randomly-selected clones, 14 contained an insert whose sequence had not been reported while the rest had an insert of either E. coli or vector origin. A search of sequence databases using the end sequences of each of the foreign inserts showed that each sequence was part of a gene encoding, in most cases, a predictable function. Our results are of significance to the collection, investigation and exploitation of the genes of uncultured microorganisms.
%K Environmental samples
%K DNA extraction
%K Mixed genomic DNA libraries
%K Uncultured microorganisms<br>环境样品
%K DNA分离纯化
%K 混合基因组DNA文库
%K 未培养微生物
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A3C6BA55AB623B90FA9104CFFC826F3C&aid=FB092188203EBB25&yid=14E7EF987E4155E6&vid=2001E0D53B7B80EC&iid=0B39A22176CE99FB&sid=76B5E24D6EC46B4B&eid=B0EBA60720995721&journal_id=0001-6209&journal_name=微生物学报&referenced_num=17&reference_num=17