Establishment of SHG-44 cell line stably and highly expressing MCHR2 / 第二军医大学学报

ABSTRACT

Objective: To construct a eukaryotic expressing vector harboring human melanin-concentrating hormone receptor 2 (MCHR2) and to establish a SHG-44 cell line stably and highly expressing MCHR2. Methods: The full-length MCHR2 cDNA fragment was amplified from the human fetal brain cDNA library by PCR and was cloned into pcDNA3.1(+) to construct eukaryotic vector pcDNA3.1(+)/MCHR2; the latter was then transduced into SHG-44 cells by Lipofectamine™. After screening culture by G418, SHG-44 cells stably expressing MCHR2 were established. The transcription and expression of MCHR2 was identified by RT-PCR, Western blotting and immunofluorescence. Results: The full-length MCHR2 cDNA fragment was amplified and the eukaryotic expression vector pcDNA3.1(+)/MCHR2 was successfully constructed. The expression of MCHR2 was found positive by RT-PCR, Western blotting and immunofluorescence, indicating that the SHG-44 cell line stably and highly expressing MCHR2 was successfully established. Conclusion: The successful establishment of MCHR2-SHG-44 cell line provides a solid foundation for further study on MCHR2 function.