载脂蛋白CⅢcDNA克隆的筛选、鉴定及其探针的制备

Screening and Identification of Apolipoprotein CⅢ cDNA Clones from Human Liver cDNA Library and Preparation of Apolipoprotein CⅢ cDNA Probe from the Clones

用载脂蛋白(apo)CⅢ抗体探针对正常人肝cDNA库进行筛选,得到4个阳性克隆。经限制性内切酶谱对其cDNA插入片段及Western blot杂交时,其表达产物的鉴定表明筛选出的4个阳性克隆为apoCⅢcDNA克隆。以此克隆制备了含apoCⅢcDNA的重组DNA,经RNA-斑点杂交分析说明,此重组DNA可用来作为apoCⅢcDNA探针。

Four of positive clones were obtainedby three runs of screening the cDNA libraryof human liver with rabbit antihumanapolipoprotein (apo) CⅢ IgG. One of themwas identified to be the positive clone ofapoCⅢcDNA by restriction endonucleasemap studids of tbe cDNA insert fragmentand by Western blot analysis of the ex-pression products. The apoCⅢcDNA-γgtllrecombinant DNA prepared from the posi-tive clone can be used in RNA--dot blotanalysis as apoCⅢ cDNA probe. And theapoCⅢcDNA clone can be also used to pro-duced apoCⅢ peptide under the induction ofisopropy1--β--D--thiogalactoside. The apoCⅢpeptide, after purification,has lot of usein further studies.