大鼠促黄体素基因β亚基的克隆和在COS细胞中的表达

Cloning and Expression of Rat LH pSubunit in COS Cell

从摘除卵巢的雌性大鼠脑垂体中提取总RBA,利用RT-PCR技术扩增了促黄体素(LH)的亚基基因。序列分析表明该基因序列与国外报道的促黄体素(LH)的亚基基因序列完全相同。将该基因片段插入到真核表达载体pcDNA3.1(-)上,构建表达质粒pcDNA3.1(-)/LH,利用脂质体包裹技术转染COS细胞,放免检测结果显示LH的表达量达6.2 miu/ml。

LH is a member of the glycoprotein honnone family which also called ICSH. The mammalian genome contains a single gene encoding the α-subunit which is common to all glycoprotein hormones. In contrast, theβ-subunit are encoded by different genes and provide the biological specificity of each hormone. We have extracted mRNA from the pituitary glands of rats, used RT-PCR to obtain theβ-subunit of LH gene and cloned it. The amplified DNA sequence is as same as the LH β-subunit gene sequence in Genebank. We have established pcDNA3.1(-)/LH plasmosome expression system and expressed the LH gene in COS cell. Those transfected cells have expressed LH βsubunit and its relative activity is 6.2miu/ml. Our final purpose is to product LH vaccine to treat BPH. Our experiment provides the possibility of the internal expression and lays the foundation of animal experiments.