Luo Jing, Lang Michael, Salzburger Walter, Siegel Nicol, Stölting Kai N, Meyer Axel
Lehrstuhl für Zoologie und Evolutionsbiologie, Department of Biology, University of Konstanz, Konstanz, Germany.
J Exp Zool B Mol Dev Evol. 2006 Nov 15;306(6):567-74. doi: 10.1002/jez.b.21112.
A goldfish (Carassius auratus auratus) bacterial artificial chromosome genomic library (BAC library) was constructed from one aquarium-bred male specimen (tetraploid, 4n=100, genome size=3.52 pg/cell). The library consists of 128,352 positive clones with an average insert size of 150.4 kb, covering the genome 11-fold. All clones were spotted onto nylon filters and thus are available for screening of genomic regions of interest, such as candidate genes, gene families, or large-sized syntenic DNA regions of cyprinid species. Preliminary screens with two genes were conducted with hybridizing probes to the genes RAG1 and lgi1. RAG1 is a single-copy gene in zebrafish and is duplicated in C. a. auratus. We found a very close correlation between the number of positive BAC clones and the expected library coverage. Two copies of lgi1 were found in zebrafish. We have detected four different copies in C. a. auratus, not in the expected abundance, which indicates some variation in the coverage of the BAC library. The preliminary screens indicate that many duplicated genes that resulted from the ancient fish-specific genome duplication persist in the tetraploid goldfish genome. Hence, the BAC library will provide a useful resource for the future work on comparative genomics, polyploidy, diploidization, and evolutionary genomics in fishes.
从一条人工养殖的雄性金鱼(四倍体,4n = 100,基因组大小 = 3.52 pg/细胞)构建了金鱼(Carassius auratus auratus)细菌人工染色体基因组文库(BAC文库)。该文库由128,352个阳性克隆组成,平均插入片段大小为150.4 kb,覆盖基因组11倍。所有克隆都点样到尼龙滤膜上,因此可用于筛选感兴趣的基因组区域,如候选基因、基因家族或鲤科鱼类的大型同线DNA区域。用两个基因进行了初步筛选,将杂交探针与RAG1和lgi1基因杂交。RAG1在斑马鱼中是单拷贝基因,在金鱼中发生了复制。我们发现阳性BAC克隆的数量与预期的文库覆盖率之间存在非常密切的相关性。在斑马鱼中发现了两个拷贝的lgi1。我们在金鱼中检测到四个不同的拷贝,但数量与预期不符,这表明BAC文库的覆盖率存在一些差异。初步筛选表明,许多由古老的鱼类特异性基因组复制产生的重复基因在四倍体金鱼基因组中仍然存在。因此,该BAC文库将为未来鱼类比较基因组学、多倍体、二倍体化和进化基因组学的研究提供有用的资源。