【摘要】 目的 为获得足量的、可供实验研究和临床应用的谷氨酸脱羧酶(GAD65)抗原。方法 将GAD65基因的cDNA正确地重组到表达型载体pGEX-3X中，并通过原核细胞进行诱导表达，SDS-聚丙烯酰胺凝胶电泳测定表达产物分子量，间接ELISA法证实其免疫原性。结果 表达产物分子量约为63.23KD，在误差允许范围(<8%)内，与GAD65蛋白分子量相近，间接ELISA显示表达产物与1型糖尿病病人血清发生明显的显色反应，证明其具有免疫原性。结论 运用基因工程技术获得的表达产物为重组GAD65抗原。这一抗原的获得，将有助于1型糖尿病的预测、预防、鉴别诊断以及免疫治疗等方面的研究。

    The expression of glutamic acid decarboxylase gene in vitro Wu Rui,Cai Jun,Zhao Chong,et al.Institute of Endocrinology,Tianjin Medical University,Tianjin,300070

    【Abstract】 Objective To get the glutamic acid decarboxylase (GAD65) antigen which can be used in experimental study and clinical application. Methods cDNA for GAD65 gene was inserted into the expression vector, and its expression in E.coli.DH5α was induced. The molecular weight of expression product was determined by SDS-PAGE and immunogenicity of expression product was ascertained by ELISA. Results The sequence analysis confirmed that GAD65 gene had been correctly inserted into pGEX-3X. The molecular weight of the expression product was approximately 63.23KD, which was within the permitted error (<8%) and similar to that of GAD65 antigen. Indirect ELISA showed that the obvious color reaction appeared after the expression products were associated with the diabetic serum. It proved that the expression products had expected immunogenicity. Conclusion The expression products obtained by the method of gene engineering are recombinant GAD65 antigen and may be used in experimental study and clinical application for diabetes mellitus.

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