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Detection， Intracellular Localization and Eukaryotic Expression In Vitro of TMEM43 Gene of Human
Zhang Haixia1,2, Li Qian3, Wang Dan4, Zhao Kexue3, Han Yumei3, Wang Xinglong3, Liang Haoqin3, Deng Yingying3, Tong Chunwei3, Li Xiangrong1, Ma Zhongren1,2, Feng Ruofei1,2*
1Key Bio-Engineering and Technology Laboratory of the Northwest Minzu University, Lanzhou 730030, China; 2Gansu Engineering Research Center for Animal Cell, Lanzhou 730030, China; 3Life Science and Engineering College of Northwest Minzu University, Lanzhou 730030, China; 4Lanzhou Minhai Bioengineering Co. Ltd, Lanzhou 730030, China
Abstract: The purpose of this research was to study the content， location， distribution， and expression of human transmembrane protein 43 (TMEM43) gene. TMEM43 gene and protein expression in MRC-5 and HEK293 cells were analyzed by RT-PCR and Western blot， respectively. TMEM43 protein distribution in MRC-5 cells was examined through indirect immunofluorescence assay (IFA). Then construction and transfection of pcDNA3.1-EGFP-TMEM43 into HEK293 cells were performed in order to detect the expression of TMEM43 gene and protein. The results showed that gene expression of TMEM43 in MRC-5 cells was significantly higher than in HEK293 cells with protein expression only occurring in MRC-5 cells. After transfecting recombinant pcDNA3.1-EGFP-TMEM43 into HEK293 cells， gene expression of TMEM43 was significantly increased along with successful protein expression. TMEM43 content of the lung cells (MRC-5) were significantly higher than the kidney cells (HEK293); the recombinant vector pcDNA3.1- EGFP-TMEM43 was successfully transfected into HEK293 cells and expression. This study provided experimental evidence for the further research into the structure， function， and related mechanisms of TMEM43.
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