Cloning of a novel mouse Gabarapl2 cDNA and its characterization  

一种新的小鼠Gabarap12 cDNA的克隆和鉴定(英文)

作　　者：陈政[1] 辛玉蓉[2] 蒋滢[1] 蒋菊香[1] 

机构地区：[1]苏州大学生命科学院,苏州215006 [2]复旦大学遗传所

出　　处：《Acta Pharmacologica Sinica》2001年第8期751-755,共5页中国药理学报（英文版）

摘　　要：AIM: To clone a novel mouse GABAA-receptor-associated protein like 2 ( Gabarapl2) gene, and to analysis its primary function. METHODS: With the aid of computer, the human GABARAPL2 cDNA was used as information probe to search mouse EST database of Gen-Bank for mouse homolog. A series of overlapping EST were found and assembled into an EST contig using Genetics Computer Group (GCG) ASSEMBLY program. The existence of the gene was then identified by experiment. Northern blotting was performed to hybridize [a-32P]dATP labeled probe with mRNA of 11 different mouse tissues that had been transferred to the nylon membrane. RESULTS: The novel gene was deposited in GenBank under Accession No AF190644. Its cDNA contained an intact open reading frame and a canonical polyadenylation signal AATAAA followed by polyA. The deduced protein was completely identical to that of human GABARAPL2, and was termed Gabarapl2 by Mouse Gene Nomenclature Committee. The putative protein of Gabarapl2 has a calculated molecular weight of 13 700 and an isoelectric point of 8.56. It was also predicted to contain two protein kinase C phosphorylation sites and one tyrosine kinase phosphorylation site. Northern hybridization showed that Gabarapl2 was expressed as a single 1.35 kb transcript, with high levels in brain, thymus, lung, heart, kidney, and liver, and low in pancreas, testis, small intestine, colon, and stomach. CONCLUSION: A novel mouse Gabarapl2 gene was cloned and identified.目的:克隆一个新的小鼠GABA_A受体相关蛋白相似蛋白2基因(Gabarapl2),并对其功能进行初步分析。方法:将已知的人GABARAPL2 cDNA序列为信息探针筛选GenBank小鼠EST数据库,将获得的一系列互相重叠的同源度较高的EST序列进行拼接。经过实验验证,在小鼠中分离和鉴定了新基因。自行制备小鼠RNA印迹膜,用杂交方法分析该基因在不同组织中的表达情况。结果:新基因在GenBank注册,登录号AF190644。同源比较显示,该推导蛋白与人GABARAPL2基因编码的蛋白完全相同,被基因命名委员会命名为小鼠Gabarapl2。该基因的推导蛋白含多个蛋白激酶磷酸化位点。杂交显示,该基因在脑、胸腺等组织表达较高,转录本大小约1.35kb。结论:克隆到一个新的小鼠Gabarapl2基因。

关 键 词：GABAA receptors HUMAN MICE amino acid sequence homology molecular models reverse transcriptase polymerase chain reaction Northern blotting DNA 

分 类 号：Q785[生物学—分子生物学]