用于转基因调控研究的人类ApoAI-AIV全基因簇克隆

Cloning of Human ApoAI-AIV Gene Cluster for Transgenic Regulation Analysis

目的 克隆人类ApoAI -AIV全基因簇用于转基因调控研究ApoAI的刺激表达药物。 方法 使用平均克隆长度为 38kb的人Cosmid基因组文库 ,通过制备逐级降低独立克隆数的文库亚库 ,以PCR对亚库中的ApoAI-AIV全基因簇首尾特征序列扩增作为亚库筛选性指标 ,筛选出独立克隆数约为 1 0数量级的文库亚库 ,进行平板单克隆菌落筛选。结果 得一个阳性克隆 ,其首尾特征序列经测序与公布序列相同。

Objective Cloning of human ApoAI-AIV gene cluster for transgenic regulation analysis.Methods Use human placenta genomic cosmid library in pWE15 vector with average 38 kb insert,made sub library pools with serial dilution of independent clones,screen positive subpools by PCR amplification of determined band covering the start ID sequence fragment and the end ID sequence fragment of ApoAI-AIV gene cluster which containing complete expression regulate element,further get positive monoclone from above subpools using plate screening method.Results Acquire 1 positive clone,which PCR amplified ID sequence was the same as genbank reported.Conclusion By subpools making and PCR amplifed ID sequence as positive indicator,we fastly got an ApoAI-AIV whole gene cluster containing positive cosmid clone for further research.