Molecular cloning of grass carp growth hormone receptor and its functional interaction with silver carp growth hormone.
A novel grass carp growth hormone receptor (gcGHR) cDNA was cloned by RT-PCR method. CHO-K1 cells were transfected with eukaryotic expression vector pcDNA3.0 containing gcGHR entire coding region, resulting in a stable CHO cell line expressing gcGHR (CHO-gcGHR). Bioactivity assay revealed that silver carp growth hormone (scGH) bound gcGHR with dissociation constant (Kd) value of 1.96 nM and stimulated CHO-gcGHR cells proliferation with the sensitivity of 20 ng/ml. The cells proliferation assay was also provided to measure fish growth hormone levels of pituitary and refolded protein.
Hu
Wei
W
Shanghai Institutes for Biological Sciences, Graduate University of the Chinese Academy of Sciences, Shanghai 200233, PR China.
Chen
Xia
X
Yu
Gusong
G
Chen
Yan
Y
Shen
Yan
Y
Zhang
Ying
Y
Gong
Yi
Y
D000818:Animals; D016466:CHO Cells; D002347:Carps / Q000378:metabolism*; D049109:Cell Proliferation; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D018076:DNA, Complementary / Q000235:genetics; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism*; D013006:Growth Hormone / Q000378:metabolism*; D011485:Protein Binding; D011986:Receptors, Somatotropin / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection
D018076:DNA, Complementary; D029941:Fish Proteins; D011986:Receptors, Somatotropin; D013006:Growth Hormone