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: To clone the cDNA of glutamic acid decarboxylase GAD65 from the brain of Wistar rats and analyze the sequence. Methods: The target gene was amplified by RTPCR. After digestion and identification, the specific DNA fragments were recombined into plasmid vector and all the nucleotide sequences were determined by dideoxy terminator method. Results: The cloned specific DNA fragment was 1758bp in length and contained 585 amino acids, including the stop codon of GAD65 full coding sequence. After repeated experiments, compared with the rat GAD65 gene provided by the EMBL nucleic acid database, it was found that the 579th base was composed of AT and a new PvuI restriction site was generated, but this change did not involve the change of amino acids. Conclusion: Obtaining the full-length cDNA of GAD65 gene of murine brain glutamate decarboxylase, which laid the foundation for the in vitro expression of this gene.