棉花学报2011，Vol.23Issue(1)：10-14,5.
雷蒙德氏棉叶绿体基因组 Fosmid 文库构建
Construction of a Fosmid Library of Chloroplast Genome in Gossypium raimondii
摘要
Abstract
In this paper, a Fosmid library of G. raimondii chloroplast genome was constructed. The chloroplast DNA was isolated by high ionic strength and low pH buffer method. The DNA was randomly sheared and cloned into pCC1FOS vector. Recombinant DNA was packaged with the Lambda Packaging Extracts, then transfected into E. coli strain EPI300. The best sheared parameter employed in the study was 18 times with middle speed using a 1 mL injector. The library of chloroplast genome (titer: l×l04 cfu·mL-1) was obtained in which the average inserted DNA fragment was 38 kb. Thirty-nine clones covering 9.2 fold the chloroplast genome were selected by selection marker to be further analyzed. Six clones, F66, F46, F28, F8, F55, and F3, which could span G. raimondii complete genome, were screened out by cotton chloroplast markers. The library would be a valuable resource for study on genome structure and functional genes investigation in cotton.关键词
雷蒙德氏棉/叶绿体DNA/Fosmid文库Key words
Gossypiurn raimondii/ cpDNA/ Fosmid library分类
生物工程引用本文复制引用
李朋波,薛龙飞,王彦霞,张曦,李召虎,华金平..雷蒙德氏棉叶绿体基因组 Fosmid 文库构建[J].棉花学报,2011,23(1):10-14,5.基金项目
教育部新世纪优秀人才支持计划(NCET-06-0106) （NCET-06-0106）
国家863计划项目(2006AA10A108,2007AA10Z125) （2006AA10A108,2007AA10Z125）
教育部长江学者和创新团队发展计划 （）
高等学校学科创新引智计划(111-2-03) （111-2-03）
