Cunningham Charles, Hikima Jun-ichi, Jenny Matthew J, Chapman Robert W, Fang Guang-Chen, Saski Chris, Lundqvist Mats L, Wing Rod A, Cupit Pauline M, Gross Paul S, Warr Greg W, Tomkins Jeff P
Marine Biomedicine and Environmental Sciences Center and Department of Biochemistry and Molecular Biology, Medical University of South Carolina, Hollings Marine Laboratory, Charleston, SC 29412, USA.
Mar Biotechnol (NY). 2006 Sep-Oct;8(5):521-33. doi: 10.1007/s10126-006-6013-9. Epub 2006 Aug 11.
Large-insert genomic bacterial artificial chromosome (BAC) libraries of two culturally and economically important oyster species, Crassostrea virginica and C. gigas, have been developed as part of an international effort to develop tools and reagents that will advance our ability to conduct genetic and genomic research. A total of 73,728 C. gigas clones with an average insert size of 152 kb were picked and arrayed representing an 11.8-fold genome coverage. A total of 55,296 clones with an average insert size of 150 kb were picked and arrayed for C. virginica, also representing an 11.8-fold genome coverage. The C. gigas and C. virginica libraries were screened with probes derived from selected oyster genes using high-density BAC colony filter arrays. The probes identified 4 to 25 clones per gene for C. virginica and 5 to 50 clones per gene for C. gigas. We conducted a preliminary analysis of genetic polymorphism represented in the C. gigas library. The results suggest that the degree of divergence among similar sequences is highly variable and concentrated in intronic regions. Evidence supporting allelic polymorphism is reported for two genes and allelic and/or locus specific polymorphism for several others. Classical inheritance studies are needed to confirm the nature of these polymorphisms. The oyster BAC libraries are publicly available to the research community on a cost-recovery basis at (www.genome.clemson.edu).
作为国际合作项目的一部分,已构建了两种在文化和经济方面具有重要意义的牡蛎物种——美国牡蛎(Crassostrea virginica)和太平洋牡蛎(C. gigas)的大插入片段基因组细菌人工染色体(BAC)文库,该项目旨在开发工具和试剂,以提高我们进行遗传和基因组研究的能力。总共挑选并排列了73,728个太平洋牡蛎克隆,平均插入片段大小为152 kb,代表了11.8倍的基因组覆盖率。对于美国牡蛎,总共挑选并排列了55,296个克隆,平均插入片段大小为150 kb,同样代表了11.8倍的基因组覆盖率。使用高密度BAC菌落滤膜阵列,用来自选定牡蛎基因的探针筛选太平洋牡蛎和美国牡蛎文库。对于美国牡蛎,每个基因探针鉴定出4至25个克隆,对于太平洋牡蛎,每个基因探针鉴定出5至50个克隆。我们对太平洋牡蛎文库中代表的遗传多态性进行了初步分析。结果表明,相似序列之间的差异程度高度可变,且集中在内含子区域。报告了两个基因支持等位基因多态性的证据,以及其他几个基因的等位基因和/或位点特异性多态性证据。需要进行经典遗传研究来确认这些多态性的性质。牡蛎BAC文库已在成本回收的基础上向研究界公开提供(网址:www.genome.clemson.edu)。