Human TSHR extracellular domain cDNA cloning and eukaryotic expression vector.docVIP

 PAGE \* MERGEFORMAT 10 Human TSHR extracellular domain cDNA cloning and eukaryotic expression vector Authors: Taketoshi, Bing-Yin, Wu Liping, Lan-ling, Wu Xiaoyan, Zhang Jin-An, Xu Li [Abstract] Objective through biotechnology to obtain thyroid-stimulating hormone receptor extracellular domain expression vector for the TSHR extracellular domain of the study provides a powerful tool. Methods from human thyroid tissue extract total RNA, RT  PCR was isolated TSHR extracellular domain cDNA, was inserted after double-digested eukaryotic expression vector pcDNA3.1 (). Results After enzyme digestion, sequencing analysis showed that the TSHR extracellular domain fragment and GenBank to provide exactly the same. Conclusion successfully constructed a human TSHR extracellular domain of eukaryotic expression vector. [Keywords:] human TSHR extracellular domain; RT  PCR; eukaryotic expression vector In autoimmune thyroid disease (autoimmune thyroid disease, AITD) in the pathogenesis of thyroid-stimulating hormone receptor (thyrotropin receptor, TSHR) is an important self-antigen [1], the body produces antibodies against the TSHR itself (thyrotropin receptor antibody, TRAb ) is the main pathogenesis. Because of the TSHR in the thyroid gland a small number of cell membrane, the structure unstable and difficult to purify TSHR unable to carry out further research. This article extracted from human thyroid RNA, obtained by reverse transcription-TSHR extracellular domain cDNA, eukaryotic expression vector to establish an effective TSHR in vitro expression system. 1 Materials and methods 1.1 Organization of the source and experimental materials Thyroid tissue from surgical patients have been diagnosed Graves. Competent DH5α bacteria were purchased in Beijing Ding States Bio-Technology Co., Ltd., pMD18  T cloning system containing TaKaRa company’s products in Japan, eukaryotic expression vector pcDNA3.1 () Department of the United States Invitrogen products. RNAgent