title: Production of biologically active recombinant goose FSH in a single chain form with a CTP linker sequence.
abstract: FSH is a glycoprotein hormone secreted by the pituitary gland that is essential for gonadal development and reproductive function. In avian reproduction study, especially in avian reproduction hormone study, it is hindered by the lack of biologically active FSH. In order to overcome this shortcoming, we prepared recombinant goose FSH as a single chain molecule and tested its biological activities in the present study. Coding sequences for mature peptides of goose FSH α and β subunits were amplified from goose pituitary cDNA. A chimeric gene containing α and β subunit sequences linked by the hCG carboxyl terminal peptide coding sequence was constructed. The recombinant gene was inserted into the pcDNA3.1-Fc eukaryotic expression vector to form pcDNA-Fc-gFSHβ-CTP-α and then transfected into 293-F cells. A recombinant, single chain goose FSH was expressed and verified by SDS-PAGE and western blot analysis, and was purified using Protein A agarose affinity and gel filtration chromatography. Biological activity analysis results showed that the recombinant, chimeric goose FSH possesses the function of stimulating estradiol secretion and cell proliferation, in cultured chicken granulosa cells. These results indicated that bioactive, recombinant goose FSH has been successfully prepared in vitro. The recombinant goose FSH will have the potential of being used as a research tool for studying avian reproductive activities, and as a standard for developing avian FSH bioassays.
authors.affiliation: Key laboratory of Animal Breeding and Reproduction, Institute of Animal Science in Jiangsu Academy of Agricultural Science, Nanjing, 210014, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Li
authors.affiliation: Key laboratory of Animal Breeding and Reproduction, Institute of Animal Science in Jiangsu Academy of Agricultural Science, Nanjing, 210014, China.
authors.forename: Huanxi
authors.initials: H
authors.lastname: Zhu
authors.affiliation: College of Animal Science, South China Agricultural University, Guangzhou, 510642, China.
authors.forename: Qinming
authors.initials: Q
authors.lastname: Qin
authors.affiliation: Key laboratory of Animal Breeding and Reproduction, Institute of Animal Science in Jiangsu Academy of Agricultural Science, Nanjing, 210014, China.
authors.forename: Mingming
authors.initials: M
authors.lastname: Lei
authors.affiliation: Key laboratory of Animal Breeding and Reproduction, Institute of Animal Science in Jiangsu Academy of Agricultural Science, Nanjing, 210014, China. zdshi@scau.edu.cn.
authors.forename: Zhendan
authors.initials: Z
authors.lastname: Shi
mesh_terms: D000818:Animals; D030161:Avian Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D006063:Chorionic Gonadotropin / Q000235:genetics*; D005260:Female; D037201:Follicle Stimulating Hormone, beta Subunit / Q000235:genetics* / Q000378:metabolism; D005777:Geese / Q000235:genetics*; D015292:Glycoprotein Hormones, alpha Subunit / Q000235:genetics* / Q000378:metabolism; D006107:Granulosa Cells / Q000166:cytology / Q000187:drug effects; D057809:HEK293 Cells; D006801:Humans; D010902:Pituitary Gland / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D030161:Avian Proteins; D006063:Chorionic Gonadotropin; D037201:Follicle Stimulating Hormone, beta Subunit; D015292:Glycoprotein Hormones, alpha Subunit; D011993:Recombinant Fusion Proteins
keywords: Bioactivity; Goose; Mammalian cell expression; Recombinant FSH; Single chain peptide