栉孔扇贝BAC文库的构建及质量评价  

Construction of Three Bacterial Artificial Chromosome(BAC) Libraries for Zhikong Scallop(Chlamys farreri)

作　　者：赵柏淞[1] 程洁[1] 陈亮[1] 于娜[1] 黄晓婷[1] 包振民[1] 

机构地区：[1]中国海洋大学海洋生物遗传学与育种教育部重点实验室,山东青岛266003

出　　处：《中国海洋大学学报（自然科学版）》2013年第11期57-63,共7页Periodical of Ocean University of China

基　　金：国家自然科学基金项目(31270047);国家高技术研究发展计划项目(2012AA10A410;2012AA092204);国家科技支撑计划项目(2011BAD13B01);现代农业产业技术体系建设专项资助

摘　　要：本研究构建了栉孔扇贝3个BAC文库，分别为BamHI文库，HindⅢ文库和Sau3AI，其中BamHI文库包含34560个克隆，平均插入片段为98kb，未发现空载；HindⅢ文库包含132480个克隆，平均插入片段为106kb，空载率为1．67％；Sau3AI文库包含23040个克隆，平均插入片段为53kb，空载率为3．33％。3个文库总共由190080个克隆组成，平均插入片段为98kb，覆盖栉孔扇贝基因组的15．4倍。通过对随机挑选BAC克隆100代的继代培养，未发现插入片段的丢失或重排现象，说明所构建的BAC文库是稳定的。将所有克隆进行超级池及二级池的构建，筛选了3个基因及遗传图谱上的7个微卫星标记，阳性克隆数在6～18之间，表明这3个文库可以在目的基因或标记的筛选、物理图谱的构建及大规模基因组的测序中发挥重要作用。We constructed three bacterial artificial chromosome （BAC） libraries for Zhikong scallop. The libraries were generated based on three restriction enzymes, BamHI, HindIII and Sau3AI, respectively. The BamHI library consisted of 34 560 clones with an average insert size of 98 kb, and none of the clones was insert free. The HindIII library consisted of 132 480 clones with an average insert size of 106 kb, and approximately 1.67 % of the clones were insert free. The Sau3AI library consisted of 23 040 clones with an average insert size of 53 kb, and approximately 3. 33% of the clones were insert free. These libraries col-lectively contained a total of 190 080 clones with an average insert size of 98 kb, covering 15.4 folds of the haploid genome. BAC stability assays showed that clones were stable during propagation. Libraries were screened with ten pairs of primer probes designed from DNA sequences of three genes and seven microsat- ellite markers. Six to 8 positive clones were identified for genes and microsatellites each. These results suggested that the 3 libraries will serve as an invaluable resource for future genetic studies on Zhikong scallop, which included gene cloning, physical mapping and even genome sequencing of this species.

关 键 词：栉孔扇贝 BAC文库 基因组 

分 类 号：S197[农业科学—农业基础科学]