草鱼腺苷酸基琥珀酸裂解酶克隆及序列分析  

MOLECULAR CLONING AND CHARACTERIZATION OF ADENYLOSUCCINATE LYASE cDNA AND GENOMIC DNA IN GRASS CARP

作　　者：屈刚[1,2] 顾继锐[1,2] 辜文博[1] 朱文漓[1] 吴江[2] 刘汉元[2] 徐恒[1,2] 

机构地区：[1]四川大学生命科学学院生物资源与生态环境教育部重点实验室,成都610064 [2]通威股份有限公司,成都610081

出　　处：《水生生物学报》2009年第6期1105-1111,共7页Acta Hydrobiologica Sinica

基　　金：通威股份公司科技发展项目-草鱼基因组计划;四川省科技攻关项目(05NG002-002-4)资助

摘　　要：腺苷酸基琥珀酸裂解酶(Adenylosuccinate lyase,ADSL)是嘌呤核苷酸合成过程中的关键酶。研究以草鱼(Ctenopharyngodon idellus)肠道cDNA文库为基础,应用PCR、RT-PCR和RACE技术,成功获得了草鱼肠道组织腺苷酸基琥珀酸裂解酶基因的cDNA全长和基因组DNA全长。该基因全长1584 bp,包含一个1449 bp的开放阅读框,编码482个氨基酸,与其他脊椎动物比对显示,其序列具有较高的保守性。草鱼腺苷酸基琥珀酸裂解酶基因组DNA由13个外显子和12个内含子组成,其外显子拼接位点非常保守,遵循GT-AG原则。Ctenopharyngodon idellus,which is common known as grass carp,is a kind of large cyprinoid fish in China.It lives in both north and south water of China and is the most important fish in Chinese fresh water aquaculture.Grass carp can efficiently transform vegetable protein into high-quality animal protein.So people call the grass carp ＂cattle and sheep in water＂.The intestine of grass carp is the main part for nutrition digestion and the functional genes from the intestinal cell are tightly connected with the efficient transformation of vegetable protein and animal protein.Adenylosuccinate lyase（ADSL） gene encoding enzymes involved in two pathways of purine nucleotide metabolism are of considerable biological and medical importance.It catalyzes two similar but separate reactions in the de novo purine biosynthesis pathway.Now,we are trying to build a connection between grass carp genome project and the industrialization of function feedstuff depended on the information that we have got from this research.In this research,we clone the full length cDNA and genomic DNA of ADSL from grass carp intestine,which is very important in the purine metabolism.The cDNA sequence of Grass Carp ADSL gene contains complete ORF starting at nucleotide 64 with a stop codon at nucleotide 1510—1512.The translated region is of 1446 nucleotides with full open reading frame（ORF） comprising 482 codons preceded by a 5′-untranslated region of 63 nucleotides and followed by 3′-untranslated region of 75 nucleotides.The typical polyadenylation signal AATAAA is found 13bp upstream of the polyA tail.The predicted mass of encoded protein is 54,552 Da with a calculated iso-electric point of 6.72 and-4.11 charges at pH 7.The ADSL amino acid sequence of grass carp has classical ＂signature＂ sequence of enzymes that catalyzing β-elimination reactions and two conservative histidine residues as general acid-base catalysis active site.The deduced amino acid sequence shares high homology with other five vertebrates ADSL and has 94.6%

关 键 词：草鱼 肠道 腺苷酸基琥珀酸裂解酶 克隆 序列分析 

分 类 号：Q781[生物学—分子生物学]