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表达序列标签(Expressed sequencetag,EST)是鉴定基因表达规律和发现新基因的一种有效的分子生物学手段。为了能在中华鲟(Acipenser sinensis Gray)中发现与生长和生殖内分泌调控相关的基因,我们构建了中华鲟垂体的SMARTcDNA质粒文库。垂体是调节生长和生殖内分泌的重要器官。在本研究中,通过测序筛选得到了944个EST克隆,将所得EST与GenBank数据库中的序列进行比对,结果表明,802(84.96%)个克隆可以找到同源序列,共代表461个基因,其中含132个已知功能基因;而142(15.04%)个克隆不能找到同源序列。研究发现,在所有基因中,阿黑皮素原基因(Proopiomelanocortin,POMC)是出现次数最高的基因,占总EST数的10.17%,显示出其在垂体中的重要地位。我们还发现了7个未知功能的基因并重点研究了其在心脏、肝脏、脾脏、肾脏、肌肉、精巢、卵巢和垂体等组织中的表达特异性。结果发现,4个基因:EG009334、EG009337、EG009338和EG009340为垂体特异性表达或垂体和卵巢特异性表达。对这些基因进一步的功能研究将有利于我们更好地了解中华鲟生长和生殖内分泌调控的分子机制。
Expressed sequence tag (EST) is an effective molecular biology method to identify the gene expression pattern and discover new genes. In order to find out the genes involved in the regulation of growth and reproductive endocrine in Acipenser sinensis Gray, we constructed a SMART cDNA plasmid library of pituitary gland of Acipenser sinensis. Pituitary is an important organ that regulates growth and reproductive endocrine. In this study, 944 EST clones were screened by sequencing and the obtained ESTs were compared with the sequences in the GenBank database. The results showed that 802 (84.96%) clones could find homologous sequences, representing a total of 461 genes, Which contains 132 known function genes; while 142 (15.04%) clones can not find homologous sequences. The study found that among all the genes, Proopiomelanocortin (POMC) was the most frequently occurring gene, accounting for 10.17% of the total ESTs, indicating its importance in the pituitary. We also identified seven genes of unknown function and focused on their expression specificity in tissues such as heart, liver, spleen, kidney, muscle, testis, ovary and pituitary. As a result, four genes, EG009334, EG009337, EG009338 and EG009340, were found to be pituitary-specific or pituitary and ovarian-specific. Further study of the function of these genes will help us to better understand the molecular mechanism of growth and reproductive endocrine regulation in Acipenser sinensis.