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Research Journals >
Indian Journal of Experimental Biology (IJEB) >
IJEB Vol.47 [2009] >
IJEB Vol.47(11) [November 2009] >
| Title: | Establishment of CHO cell line expressing human MCHR2 gene and research of its molecular characteristics |
| Authors: | Yuan, Chengfu
Bu, Youquan
Sengupta, Joyeeta
Yang, Junxia
Huang, Xiuning
Cheng, Li
Zhang, Qin
Yi, Faping
Liu, Geli
Song, Fangzhou |
| Keywords: | Eukaryotic expression vector
Gene expression
MCHR2
Molecular characteristics |
| Issue Date: | Nov-2009 |
| Publisher: | CSIR |
| Abstract: | The whole
length of MCHR2 gene cDNA fragment was amplified by PCR using human fetal brain
cDNA library as template. The pcDNA3.1
(+)/MCHR2 eukaryotic expression vector was constructed successfully. The recombinant
pcDNA3.1 (+)/MCHR2 plasmid was transfected into Chinese hamster ovary (CHO)
cell by lipofectamineTM2000, after G418 selection and then the CHO
cell line expressing MCHR2 gene was established. The MCHR2 gene expression was
tested by RT-PCR, western blotting and immunofluorescence. The maximum binding (Bmax)
of CHO cell line was 309.97±1.14 fM·mg-1protein and the dissociation constant (Kd value)
was 0.170±0.0006 nM. MCH could
stimulate Ca2+ release, its 50% effective concentration (EC50)
was 2.32±0.01 nM. The construction of the CHO cell line
and the research of MCHR2 molecular characteristics have established a good
experimental basis for the further research about the function of MCHR2 gene. |
| Page(s): | 862-870 |
| ISSN: | 0975-1009 (Online); 0019-5189 (Print) |
| Source: | IJEB Vol.47(11) [November 2009]
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