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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

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【作者】 沈琪汪兴生沈子龙吴国球

【Author】 SHEN Qi, WANG Xing-sheng ,SHEN Zi-long ,WU Guo-qiu(1. China Pharmaceutical University;2. College of Medcine, Southeneast University, Nanjing 210009)

【机构】 中国药科大学生物技术中心东南大学医学院检验科 南京210009

【摘要】 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。

【Abstract】 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.

【关键词】 促黄体素基因克隆基因表达
【Key words】 LHGene clonePlasmosome expression
  • 【文献出处】 药物生物技术 ,Pharmaceutical Biotechnology , 编辑部邮箱 ,2003年01期
  • 【分类号】Q786
  • 【被引频次】6
  • 【下载频次】78
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