Gene Cloning, Expression And Physiological Activity Of Ghrelin In Guizhou Zhuxiang Black-Feather Chicken

Ghrelin is the endogenousli gand for the growth hormone secretagague receptor and at present,except that the growth hormone release hormone and somatostatin, endogenous material that the third regulation GH secreted.Ghrelin of the mammals could promote the growth hormone to release and raise the appetite of animals.It plays an important role to balance to regulate energy,food intake and other physiology process.And it becomes a major braingut peptide in recent years.At present the study about Ghrelin in animal is less,In the Guizhou Zhuxiang Black-Feather Chicken,the relationship is unclear between Ghrelin and its growth and feeding of regulation.In this research,we studied Ghrelin gene structure,expression and physiological activity by PCR,RT-PCR techniques and others to obtain Ghrelin DNA,Ghrelin cDNA.And succeeded to construction Ghrelin prokaryotic expression plasmid.The recombinant plasmid PTYB11/G were obtained.By the optimization, Definited the best system of expression conditions is 5 hours induction,37℃, OD₆₀₀=0.5 in E.coil.ER2566.In these conditions,the expression of fusion protein of is 40%of total protein.By the affinity chromatography,fusion protein is purificated. The antiserum of chicken and rabbit were prepared with the fusion protein and Ghrelin-KLH.With active immunity and passive immunity,confirmed that Ghrelin of chicken have no obviously impact on ultimum metabolite,plasma Insulin,glucagons, T₃ and T₄.It entrances plasma GH,but inhibit food intake and increase of fat and body weight.These conclusion are obviously different from the mammal.The results make foundation with scale cultivation of Guizhou Zhuxiang Black-Feather Chicken.