摘要
细菌人工染色体(BAC)文库是进行生物基因组学研究的一个重要手段。以家蚕微孢子虫(Nosema bomby-cis)重庆分离株为材料,在前期构建的家蚕微孢子虫基因组框架图数据基础上,对构建家蚕微孢子虫BAC文库过程中的关键技术,如脉冲胶块(plug)的制备与处理、限制性酶的筛选、DNA的部分消化、酶切片段的选择、插入DNA片段与载体的摩尔比值等进行了研究,建立了构建家蚕微孢子虫BAC文库适宜的技术体系。构建的家蚕微孢子虫BAC文库由6 478个克隆构成,克隆片段平均大小约为50 kb,相当于家蚕微孢子虫基因组(15.33 Mb)的20倍,文库空载率较低,有利于深入开展家蚕微孢子虫基因组相关方面的研究。
Bacterial artificial chromosome（BAC） is one of the important means to genomic research of organisms. Based on the database of draft sequences of Nosema bombycis（Chongqing isolate） genome, key processes in constructing the BAC library, such as preparation of plugs, selection of enzymes, partial digestion of DNA fragments, selection of digested fragments, molecular ratio of inserted DNA to vector DNA, were explored. As a result, a BAC library of the Nosema bombycis has been constructed with more than 6 478 clones based upon the optimized method. The mean size of inserted fragments is -50 kb, indicating a nearly 20-fold coverage of the Nosema bombycJs haploid genome and a very low rate of empty vectors. The constructed library establishes a good foundation for further analysis on Nosema bombycis genome.
出处
《蚕业科学》
CAS
CSCD
北大核心
2009年第2期314-319,共6页
ACTA SERICOLOGICA SINICA
基金
重庆市自然科学基金重大项目(编号CSTC2006AA5019)
国家茧丝办风险基金项目(编号M012005-000Y-00070)
国家重点基础研究发展计划“973”项目(编号2005CB12-1000)
关键词
家蚕微孢子虫
基因组
细菌人工染色体文库
Noserna bombycis
Genome
Bacterial artificial chromosome（BAC） Library
