[Expression of grass carp growth hormone in the yeast Pichia pastoris].
Growth hormone plays an essential role in the stimulation of somatic growth and development. To produce large amount of grass carp growth hormone (gcGH) of biological activity for further use, this study aimed at high-yield expression of gcGH in the yeast Pichia pastoris. The gcGH cDNA was inserted into a yeast vector, pGAPZ-alpha-B. Under the control of the promoter GAP (glyceraldehyde 3-phosphate dehydrogenase), a 22-kilodalton protein, similar to the molecular weight of native growth hormone, was detected in the supernatant of transformed yeast. It led to a level as high as of 50 mg/L. This protein could be specifically immunoprecipitated with rabbit polyclonal antibodies raised against gcGH. A receptor ELISA Sandwich method was also performed to verify the biological activity of the in vitro expressed gcGH.
Wang
Wei
W
State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan 430072, China.
Sun
Yong-Hua
YH
Wang
Ya-Ping
YP
Zhu
Zuo-Yan
ZY
D000818:Animals; D001667:Binding, Competitive; D015153:Blotting, Western; D002347:Carps / Q000378:metabolism*; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D013006:Growth Hormone / Q000235:genetics / Q000378:metabolism*; D010843:Pichia / Q000235:genetics*; D011986:Receptors, Somatotropin / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D011986:Receptors, Somatotropin; D011994:Recombinant Proteins; D013006:Growth Hormone