自身抗原核点蛋白Sp100基因克隆和真核表达
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王雪松, 李永哲, 刘国振
原发性胆汁性肝硬化;核点蛋白Sp100基因;自身抗原;克隆王雪松,李永哲,刘国振.自身抗原核点蛋白Sp100基因克隆和真核表达. 世界华人消化杂志2006;14(8)758-762,自身抗
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李永哲, 王雪松, 中国医学科学院中国协和医科大学、北京协和医院检验科, 北京市 100730
刘国振, 中国科学院北京华大基因研究中心 北京市 101300
王雪松, 女, 1979-06-23生, 吉林省吉林市人, 汉族, 2006年北华大学医学院毕业, 在读硕士, 主要从事自身免疫性疾病研究.国家自然科学基金项目, No. 30471617
国家高技术研究发展计划(863计划)重大专项基金资助项目, No. 2002AARZ2011
通讯作者: 李永哲, 100730, 北京市, 中国医学科学院中国协和医科大学、北京协和医院检验科. yongzhelipumch@yahoo.com.cn
电话: 010-65295416 传真: 010-65295416
收稿日期: 2006-01-11 接受日期: 2006-02-08
Gene cloning and fusion expression of human autoantigen Sp100 in yeasts
Xue-Song Wang, Yong-Zhe Li, Guo-Zhen Liu
Yong-Zhe Li, Xue-Song Wang, Department of Laboratory, Hospital of Peking Union Medical College, Beijing 100730, China
Guo-Zhen Liu, Beijing Huada Genomics Research Center, Chinese Academy of Sciences, Beijing 101300, China
Supported by National Natural Science Foundation of China, No. 30471617, and the Key Special Foundation of National 863 Project, No. 2002AARZ2011
Correspondence to: Yong-Zhe Li, Department of Laboratory, Hospital of Peking Union Medical College, Beijing 100730, China. yongzhelipumch@yahoo.com.cn
Received: 2006-01-11 Accepted: 2006-02-08
Abstract
AIM: To clone and construct the plasmid cont-aining human autoantigene Sp100 gene, and then to identify the immunoreactivity of the purified recombinant protein.
METHODS: The Sp100 gene was amplified from human liver cDNA library, and then was cloned into PEGH vector to induce the Sp100 expression. The obtained products were identified and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot.
RESULTS: The sequence of Sp100 autoantigene gene was confirmed to be the same as the sequence reported in GenBank. The fusion pro-teins were found at 53-, 55-, 52-, 37-, 42-, and 47-ku strip on SDS-PAGE gel. Western blot analysis showed that the fusion protein with 55, 52, 42 and 47 ku had the same immunoreactivity as human Sp100 autoantigene ......
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