Abstract:
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 A cquire 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.