Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, 4-5-7 Konan, Minato-ku, Tokyo, 108-8477, Japan.
BMC Genomics. 2010 Feb 26;11:141. doi: 10.1186/1471-2164-11-141.
Higher crustaceans (class Malacostraca) represent the most species-rich and morphologically diverse group of non-insect arthropods and many of its members are commercially important. Although the crustacean DNA sequence information is growing exponentially, little is known about the genome organization of Malacostraca. Here, we constructed a bacterial artificial chromosome (BAC) library and performed BAC-end sequencing to provide genomic information for kuruma shrimp (Marsupenaeus japonicus), one of the most widely cultured species among crustaceans, and found the presence of a redundant sequence in the BAC library. We examined the BAC clone that includes the redundant sequence to further analyze its length, copy number and location in the kuruma shrimp genome.
Mj024A04 BAC clone, which includes one redundant sequence, contained 27 putative genes and seemed to display a normal genomic DNA structure. Notably, of the putative genes, 3 genes encode homologous proteins to the inhibitor of apoptosis protein and 7 genes encode homologous proteins to white spot syndrome virus, a virulent pathogen known to affect crustaceans. Colony hybridization and PCR analysis of 381 BAC clones showed that almost half of the BAC clones maintain DNA segments whose sequences are homologous to the representative BAC clone Mj024A04. The Mj024A04 partial sequence was detected multiple times in the kuruma shrimp nuclear genome with a calculated copy number of at least 100. Microsatellites based BAC genotyping clearly showed that Mj024A04 homologous sequences were cloned from at least 48 different chromosomal loci. The absence of micro-syntenic relationships with the available genomic sequences of Daphnia and Drosophila suggests the uniqueness of these fragments in kuruma shrimp from current arthropod genome sequences.
Our results demonstrate that hyper-expansion of large DNA segments took place in the kuruma shrimp genome. Although we analyzed only a part of the duplicated DNA segments, our result suggested that it is difficult to analyze the shrimp genome following normal analytical methodology. Hence, it is necessary to avoid repetitive sequence (such as segmental duplications) when studying the other unique structures in the shrimp genome.
较高等甲壳动物(甲壳纲 Malacostraca 类)是最具物种多样性和形态多样性的非昆虫节肢动物群体,其许多成员具有商业价值。尽管甲壳动物的 DNA 序列信息呈指数级增长,但对甲壳纲动物的基因组组织知之甚少。在这里,我们构建了一个细菌人工染色体(BAC)文库,并进行了 BAC 末端测序,为甲壳动物中最广泛养殖的物种之一——日本对虾(Marsupenaeus japonicus)提供了基因组信息,并发现 BAC 文库中存在冗余序列。我们检查了包含冗余序列的 BAC 克隆,以进一步分析其在日本对虾基因组中的长度、拷贝数和位置。
包含一个冗余序列的 Mj024A04 BAC 克隆包含 27 个假定基因,似乎显示出正常的基因组 DNA 结构。值得注意的是,在假定基因中,有 3 个基因编码凋亡抑制蛋白同源蛋白,有 7 个基因编码与白斑综合征病毒同源蛋白,后者是一种已知影响甲壳类动物的致病病原体。381 个 BAC 克隆的菌落杂交和 PCR 分析表明,近一半的 BAC 克隆保持着与其代表性 BAC 克隆 Mj024A04 同源的 DNA 片段。在日本对虾核基因组中,至少以 100 个拷贝数检测到 Mj024A04 的部分序列。基于微卫星的 BAC 基因分型清楚地表明,来自至少 48 个不同染色体位点克隆了 Mj024A04 同源序列。与已有的 Daphnia 和 Drosophila 基因组序列缺乏微同线性关系表明,这些片段在当前节肢动物基因组序列中是日本对虾所特有的。
我们的结果表明,日本对虾基因组中发生了大量 DNA 片段的超扩张。尽管我们只分析了部分重复的 DNA 片段,但我们的结果表明,按照常规分析方法研究虾基因组是很困难的。因此,在研究虾基因组的其他独特结构时,需要避免重复序列(如片段重复)。