蛋白酶体β5亚单位基因真核表达载体的构建

Construction of the recombinant expression plasmid encoding proteasome β5 subunit gene

目的:构建蛋白酶体亚单位β5基因真核表达质粒。方法:从人晶状体上皮细胞株SRA01/04中提取总RNA,经RT-PCR扩增获得β5亚单位的全长cDNA片段,将其克隆至真核表达载体pcDNA3.1上。结果:RT-PCR法扩增出约792bp的β5亚单位基因全部编码序列的片段。酶切鉴定和测序分析证实所插入的β5亚单位的基因序列完全正确。结论:成功构建了蛋白酶体β5亚单位基因真核表达重组质粒。

AIM： To clone and construct an eukaryotic expression plasmid containing β5 subunit gene, in order to study the mechanism of age-related cataract formation and the prevention measure. ·METHODS： Total RNA was extracted from human lens epithelium strain SRA01/04. β5 subunit cDNA was ampli-fied by RT-PCR, ligated into the eukaryotic expression vector pcDNA3.1（＋）.·RESULTS： RT-PCR product is about 792bp specific segement. Analysis by estricting enzyme EcoRⅠand HindⅢ and DNA sequence showed the inserted β5 subunit gene was correct.·CONCLUSION： Eukaryotic expression plasimd pcDNA 3.1-β5 is successfully constructed in order to make a basis for the further reseach.