家鸡TSHβ、FSHβ和LHβ重组蛋白表达及纯化研究

Expression and Purification of Recombinant Chicken TSHβ, FSHβ, and LHβ Proteins

摘要:

促性腺激素（促卵泡激素FSH、黄体生成素LH）和促甲状腺激素（TSH）都是由垂体分泌的糖蛋白类激素.促性腺激素在脊椎动物的性腺发育和功能调节中起关键作用.促甲状腺激素通过刺激甲状腺激素（T4、T3）的分泌从而参与脊椎动物的生长发育、代谢等生理过程调节.本文中利用RT-PCR、分子克隆等技术，构建了家鸡FSHβ亚基、LHβ亚基和TSHβ亚基的原核表达质粒, 在细菌中高效表达了TSHβ亚基、FSHβ亚基和LHβ亚基的重组蛋白；利用离子亲和层析蛋白纯化技术获得了高质量的重组蛋白. 这为后续制备三种蛋白抗体奠定基础.

Abstract:

Gonadotropin（Follicle-stimulating hormone (FSH), Luteinizing hormone (LH)）and Thyroid-stimulating hormone (TSH) are glycoprotein secreted by pituitary. Gonadotropin plays a key role in the regulation of the development and function of vertebrate gonads. TSH participates in development and metabolism by stimulate the thyroid to secrete T4 and T3. In the study, we prepared the prokaryotic expression plasmids encoding chicken TSHβ subunit, FSHβ subunit and LHβ subunit and produced the three recombinant proteins using RT-PCR and molecular approaches. We also purified the recombinant proteins by Metal Ion Affinity Chromatography. This work paves the way for future preparation of polyclonal antibodies against chicken TSHβ, FSHβ and LHβ in rabbits.