摘要
3-磷酸甘油醛脱氢酶(glyceraldehyde-3-phosphatede hydrogenase,GAPDH)是维持生命最基本活动的关键酶之一。采用RT-PCR、RACE等技术,获得了拟穴青蟹gapdh基因全长cDNA序列。该序列全长1 440 bp,开放阅读框(ORF)为1 008 bp,编码335个氨基酸残基。同源分析显示,该基因编码的蛋白与其他一些物种具有很高相似性,推测gapdh基因具有很高的保守性。经荧光定量PCR检测,gapdh基因在拟穴青蟹多个组织中均有表达,且在胸神经团、眼柄神经节、卵巢、表皮中表达量较高。在拟穴青蟹卵巢发育过程中,gapdh基因在卵巢发育早期(Ⅱ期)表达量最高,在卵巢发育成熟期(Ⅴ期)表达量最低,由此推测GAPDH主要参与了卵巢的细胞分裂增殖过程。
Glyceraldehyde-3-phosphatede hydrogenase（GAPDH）is one of the basic enzymes that maintain the life activities.In this paper,gapdh gene was isolated from the mud crab,Scylla paramamosain using RT-PCR and RACE methods.The obtained full-length cDNA of GAPDH was 1 440 bp with an open reading frame of 1 008 bp encoding a putative peptide of 335 amino acids.By alignment,the amino acid sequence of S.paramamosain GAPDH showed high homology with those of some other animals.It suggested gapdh was highly conservative.Real-time PCR showed that the gapdh gene was expressed in various tissues,and highly expressed in thoracic ganglia,eyestalk ganglia,ovary and epidermis.During the ovarian development,the expression of gapdh mRNA was significantly higher at early-developing stage（Ⅱstage）and lowest at ripe stage（Ⅴstage）.It indicated that GAPDH might play an important role in cell mitosis in ovary of the mud crab.
出处
《水产学报》
CAS
CSCD
北大核心
2013年第1期55-62,共8页
Journal of Fisheries of China
基金
国家自然科学基金项目(40406030
41076081
31272632)
