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目的 为应用基因工程技术生产适用于 1型糖尿病早期诊断的试剂盒打下基础。方法 从白种人胰腺细胞和中国人胰岛细胞瘤组织总 RNA中 ,经逆转录 -聚合酶链反应 (RT- PCR)克隆了胰岛细胞自身抗原 6 9k D蛋白 (ICA6 9)基因的 c DNA,双脱氧末端终止法对其全部核苷酸序列予以确定后 ,将此编码 483个氨基酸、全长 144 9bp的 c DNA重组入表达型质粒中。结果 核苷酸序列分析证实重组质粒接口处读码框架正确。结论 此构建为 ICA6 9重组基因的表达及表达产物在临床诊断中的应用奠定了基础。
Objective To lay the foundation for the production of kits for early diagnosis of type 1 diabetes using genetic engineering techniques. Methods The c DNA of islet cell autoantigen 6 9k D (ICA6 9) gene was cloned by RT-PCR from total RNA of human pancreatic cells and Chinese islet cell tumor tissues. After all the nucleotide sequences were confirmed by dideoxy terminator method, the c DNA of 489 amino acids and a total length of 144 9bp was recombined into the expression plasmid. Results Nucleotide sequence analysis confirmed that the reading frame at the recombination plasmid interface was correct. Conclusion This construction laid the foundation for the expression of ICA6 9 recombinant gene and its application in clinical diagnosis.