| ICA69基因工程融合抗原的制备 |
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| 引用本文： | 黄鹤,甘一如,黄乃萍,张镜宇.ICA69基因工程融合抗原的制备[J].天津大学学报(英文版),2002,8(4):231-234. |
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| 作者姓名： | 黄鹤 甘一如 黄乃萍 张镜宇 |
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| 作者单位： | 1. 天津大学化工学院,天津,300072
2. 天津医科大学内分泌研究所,天津,300072 |
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| 摘 要： | 基于基因工程技术 ,用PCR法扩增出编码ICA6 9的cDNA片段 ,直接克隆到pSPORT 1质粒上 ,经DNA序列测定 ,插入到GST融合蛋白表达载体 pGEX 2T ,构成重组质粒 p2T ICA6 9,得到的表达产物GST ICA6 9融合蛋白用间接ELISA法检测其免疫原性 .测序结果表明 ,所获PCR产物已正确重组到PGEX 2T表达型质粒中 .重组质粒在原核细胞中表达的融合蛋白具有免疫原性 ,并能应用于Ⅰ型糖尿病病人血清中抗ICA6 9抗体的检测 .所获得的表达产物为重组ICA6 9融合抗原 ,有助于提高Ⅰ型糖尿病的预报率和确诊率 .
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| 关 键 词： | 胰岛细胞自身抗原 69kD蛋白 GST融合蛋白 Ⅰ型糖尿病 酶联免疫吸附试验 |
Preparation of Recombinant Islet Cell Autoantigen 69 kD Fusion Protein |
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| HUANG He,GAN Yi-ru,HUANG Nai-ping,ZHANG Jing-yu.Preparation of Recombinant Islet Cell Autoantigen 69 kD Fusion Protein[J].Transactions of Tianjin University,2002,8(4):231-234. |
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| Authors: | HUANG He GAN Yi-ru HUANG Nai-ping ZHANG Jing-yu |
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| Abstract: | To get recombinant antigen ls l et Cell Autoantigen 69 ( ICA69 ) which was expressed in Escherichiacoli strains (E. coli) by means of the gene engineerirg technique so that it can be used for early diagnosis ofand screening in type Ⅰ diabetes mellitus, the cDNA fragment of human ICA69 was amplified by PCR, andthen cloned into pSPORT 1 vector. After DNA sequencing, it was inserted into pGEX-2T between the sites ofEcoR I and Sma I , then recombinant plasmid p2T-ICA69 was constructed and introduced into E. coli. TheGST-ICA69 fusion protein was expressed by the induction of IPTG. The recombinant ICA69 proteins were usedto detect the antibodies against hICA69 in 100 healthy subjects and type Ⅰ diabetic serum by the use of indirectELISA. The sequence analysis showed that the amplified fragments contained 1449 bp, encoded 483 aminoacids, and had been correctly inserted into pGEX-2T vector. The recombinant proteins expressed in the pro-karyotic cells had immunogenicity and could be used to detect antibodies against ICA69 in type Ⅰ diabetic ser-um. Finally it can be concluded in this paper that the expression products obtained by the method of gene engi-neering are recombinant ICA69 antigen and may be used to improve the forecast rate and the diagnostic rate oftype Ⅰ diabetes in combination with other tests. |
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| Keywords: | islet cell autoantigen 69 kD protein GST fusion protein IDDM ELISA |
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