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| 人IA-2基因的部分区段克隆表达及其在1型糖尿病诊断中的应用价值评估 | |
| 引用本文： | 王国华,张贺秋,宋晓国,陈坤,朱翠侠,楚晓燕,刘喜明,戴振华,方平,冯晓燕. 人IA-2基因的部分区段克隆表达及其在1型糖尿病诊断中的应用价值评估[J]. 生物技术通讯, 2010, 21(1): 12-16. DOI: 10.3969/j.issn.1009-0002.2010.01.003 |
| 作者姓名： | 王国华 张贺秋 宋晓国 陈坤 朱翠侠 楚晓燕 刘喜明 戴振华 方平 冯晓燕 |
| 作者单位： | 1. 军事医学科学院基础医学研究所,北京,100850 2. 中国中医科学院广安门医院,北京,100053 3. 上海海泰金芯生物分子检测技术有限公司,上海,201203 |
| 摘 要： | 目的:构建人IA-2基因不同区段原核表达载体,诱导表达获得重组蛋白,并初步验证其在1型糖尿病蛋白酪氨酸磷酸酶自身抗体检测中的价值。方法:用RT-PCR方法调取目的基因,构建相应的原核表达质粒,转化大肠杆菌HB101,诱导表达获得纯化重组蛋白;以重组蛋白为包被抗原,初步建立检测蛋白酪氨酸磷酸酶自身抗体的ELISA方法,评价各片段在1型糖尿病诊断中的价值。结果:获得了2种可被1型糖尿病患者血清识别的重组人蛋白酪氨酸磷酸酶抗原区段IA-2(601～979)和IA-2(683～979),检测敏感性和特异性相当,但IA-2(683～979)检测的阳性D450nm值明显高于IA-2(601～979),成为首选的抗原区段。结论:所选重组人IA-2(683～979)抗原区段具有良好的抗原性,可作为1型糖尿病患者辅助诊断试剂的候选抗原。 |
| 关 键 词： | 1型糖尿病 蛋白酪氨酸磷酸酶 蛋白酪氨酸磷酸酶自身抗体 酶联免疫吸附测定法 |
Cloning,Expression and Diagnostic Evaluation of the Fragments of Human Protein Tyrosine Phosphatase-Like Protein IA-2 |
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| WANG Guo-Hua,ZHANG He-Qiu,SONG Xiao-Guo,CHEN Kun,ZHU Cui-Xia,CHU Xiao-Yan,LIU Xi-Ming,DAI Zhen-Hua,FANG Ping,FENG Xiao-Yan. Cloning,Expression and Diagnostic Evaluation of the Fragments of Human Protein Tyrosine Phosphatase-Like Protein IA-2[J]. Letters in Biotechnology, 2010, 21(1): 12-16. DOI: 10.3969/j.issn.1009-0002.2010.01.003 | |
| Authors: | WANG Guo-Hua ZHANG He-Qiu SONG Xiao-Guo CHEN Kun ZHU Cui-Xia CHU Xiao-Yan LIU Xi-Ming DAI Zhen-Hua FANG Ping FENG Xiao-Yan |
| Affiliation: | WANG Guo-Hua1,ZHANG He-Qiu1,SONG Xiao-Guo1,CHEN Kun1,ZHU Cui-Xia1,CHU Xiao-Yan2,LIU Xi-Ming2,DAI Zhen-Hua1,FANG Ping3,FENG Xiao-Yan1 1.Beijing Institute of Basic Medical Sciences,Beijing 100850,2.Guang'anmen Hospital,China Academy of Chinese Medical Sciences,Beijing 100053,3.Shanghai Haitaijinxin Biomolecule Detection Technology Co.,Ltd,Shanghai 201203,China |
| Abstract: | Objective:To obtain two different recombinant fragments of human protein tyrosine phosphatase-like protein IA-2,and to evaluate their diagnostic application for type 1 diabetes mellitus.Methods:The two DNA fragments of the IA-2 were obtained by RT-PCR and nested-PCR.The corresponding prokaryotic expression vector pBVIL1/IA-2 was constructed and transformed into E.coli HB101 to induce the expression.The two antigen fragments were used as the coating antigens,and the ELISA was established for the detection of... |
| Keywords: | type 1 diabetes mellitus recombinant human protein tyrosine phosphatase-like protein IA-2 IA-2 autoantibody enzyme-linked immunosorbent assay |
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