Construction of eukaryotic expressing vector of human melanin-concentrating hormone receptor 1 and establishment of stable transfectant CHO cell line
- VernacularTitle:人黑色素浓集激素1型受体真核表达载体的构建及稳定转染CHO细胞系的建立
- Author:
Chengfu YUAN
;
Junxia YANG
;
Lili WEI
- Publication Type:Journal Article
- Keywords:
receptor, melanin-concentrating hormone, type 1;
eukaryotic expression vector;
CHO cells;
gene expression;
transfection
- From:
Medical Journal of Chinese People's Liberation Army
1982;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct eukaryotic expressing vector of human melanin-concentrating hormone receptor 1(MCHR1), then to transfect CHO cells with the vector for establishment of stable CHO cell line. Methods The full-length MCHR1 cDNA fragment was amplified by PCR from the human fetal brain cDNA library and then inserted into eukaryotic expression vector pcDNA3.1(+).The recombinant was transfected into CHO cells by lipofectamine TM 2000 after identification of digestion and sequencing on the recombinant eukaryotic expression vector pcDNA3.1(+)/MCHR1. The stable transfected CHO cell line was then established by screening cultures with G418, and the transcription and expression of MCHR1 were identified by RT-PCR, Western blot and immunofluorescence. Results The eukaryotic expression vector pcDNA3.1(+)/MCHR1 was constructed successfully, stable transfected CHO cell line was established, the MCHR1 protein was expressed successfully. Conclusion The construction of eukaryotic expression vector pcDNA3.1(+)/MCHR1 and the establishment of stable transfected CHO cell line provided a solid experimental foundation for further studies on the function of MCHR1.
