Jiangsu Key Laboratory for Biodiversity and Biotechnology, College of Life Sciences, Nanjing Normal University, Nanjing 210046, China.
Biochem Genet. 2011 Oct;49(9-10):645-55. doi: 10.1007/s10528-011-9439-3. Epub 2011 May 24.
Expressed sequence tags (ESTs) obtained from two-directional suppression subtractive hybridization (SSH) of cDNA libraries derived from the pleon of the Chinese mitten crab were analyzed using mRNA subtractive hybridization. After cDNA cloning and sequencing, gene expression profiles were examined, focusing on brachyurization. We assembled 211 non-redundant ESTs from SSH library 1 (yielding 130 unique transcripts: 25 consensi, 105 singletons) and 669 from SSH library 2 (195: 51 consensi, 144 singletons). Functional analysis shows that these genes are putatively involved in various cellular processes (33%), ribosomal RNA/proteins (30%), molting (12%), signal transduction (6%), immune factor and stress proteins (6%), development (4%), energy metabolism (3%), and mitochondrial membrane protein, hormone metabolizing, and chaperone (1% each) of other organisms. Some 3% are of unknown function in SSH library 1. The results facilitate the functional study of candidate genes involved in early developmental processes, especially the regulation of brachyurization in this crab.
采用 mRNA 消减杂交技术对双向抑制消减杂交(SSH) cDNA 文库获得的中华绒螯蟹腹部 cDNA 进行了表达序列标签(EST)分析。在 cDNA 克隆和测序后,我们对基因表达谱进行了研究,重点关注短腕幼虫化。我们从 SSH 文库 1 组装了 211 个非冗余 EST(产生 130 个独特转录本:25 个共识,105 个单核苷酸),从 SSH 文库 2 组装了 669 个 EST(195 个:51 个共识,144 个单核苷酸)。功能分析表明,这些基因可能参与各种细胞过程(33%)、核糖体 RNA/蛋白质(30%)、蜕皮(12%)、信号转导(6%)、免疫因子和应激蛋白(6%)、发育(4%)、能量代谢(3%)以及其他生物体的线粒体膜蛋白、激素代谢和伴侣(各占 1%)。SSH 文库 1 中约有 3%的基因功能未知。这些结果有助于研究参与早期发育过程的候选基因的功能,特别是调控这种螃蟹短腕幼虫化的基因。