Production of biologically active recombinant goose FSH in a single chain form with a CTP linker sequence
文献类型： 外文期刊
作者： Li, Hui 1 ; Zhu, Huanxi 1 ; Qin, Qinming 2 ; Lei, Mingming 1 ; Shi, Zhendan 1 ;
作者机构： 1.Jiangsu Acad Agr Sci, Key Lab Anim Breeding & Reprod, Inst Anim Sci, Nanjing 210014, Jiangsu, Peoples R China
2.South China Agr Univ, Coll Anim Sci, Guangzhou 510642, Guangdong, Peoples R China
关键词： Goose;Recombinant FSH;Single chain peptide;Mammalian cell expression;Bioactivity
期刊名称：MOLECULAR BIOLOGY REPORTS （ 影响因子：2.316； 五年影响因子：2.357 ）
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收录情况： SCI
摘要： 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 alpha and beta subunits were amplified from goose pituitary cDNA. A chimeric gene containing alpha and beta 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-gFSH beta-CTP-alpha 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.
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