鼠脑谷氨酸脱羧酶GAD_(65)基因的cDNA克隆  

cDNA CLONING AND SEQUENCING OF GLUTAMIC ACID DECARBOXYLASE(GAD 65 ) FROM RAT BRAIN

作　　者：蔡军[1] 吴锐[1] 赵崇[1] 何炜 张镜宇[1] 

机构地区：[1]天津医科大学内分泌研究所

出　　处：《天津医科大学学报》1998年第4期311-313,共3页Journal of Tianjin Medical University

基　　金：卫生部科学基金

摘　　要：目的：从Ｗｉｓｔａｒ大鼠脑中克隆谷氨酸脱羧酶ＧＡＤ６５的ｃＤＮＡ并进行序列分析。方法：用ＲＴＰＣＲ法扩增目的基因，酶切鉴定后，将特异性ＤＮＡ片段重组入质粒载体中，双脱氧末端终止法测定其全部核苷酸顺序。结果：克隆的特异性ＤＮＡ片段为编码５８５个氨基酸、含终止密码子在内的共１７５８ｂｐ的ＧＡＤ６５全编码序列。经重复实验，与ＥＭＢＬ核酸数据库提供的大鼠ＧＡＤ６５基因比较，发现第５７９位碱基由ＡＴ，并产生一新的ＰｖｕＩ酶切位点，但这种变化不涉及氨基酸的改变。结论：获得鼠脑谷氨酸脱羧酶ＧＡＤ６５基因的全长ｃＤＮＡ，为该基因的体外表达打下基础。Objective: cDNA cloning and sequencing of glutamic acid decarboxylase(GAD 65 ) from rat brain. Methods: The full-length cDNA of GAD 65 containing 1758bp coding for 585 amino acids was amplified by use of RT PCR and finally inserted into pUC vector system for cloning. Then, its nucleotide sequence was determined by the dideoxy chain termination method combined with PCR technique. Results: Only the 579th base is different from that of provided by EMBL nucleotide data bank(AT), which produces another Pvu11 restriction site. It does not involve in changing the corresponding amino acid. Conclusion: The rat full length cDNA of GAD 65 was obtained, which was an preliminary work for its expression in vitro.

关 键 词：谷氨酸脱羧 CDNA 克隆 RT-PCR 糖尿病 

分 类 号：Q554.1[生物学—生物化学] R587.104[医药卫生—内分泌]