Life Sciences |
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CONSTRUCTION AND IDENTIFICATION OF THE RECOMBINANT OF THE AAV VECTOR AND HUMAN INTERFERON-GAMMA |
ZHU Hai-hong, CHEN Zhi, ZHANG Ming-tai, DING Lie-ming |
Institute of Infectious Diseases, College of Midicine, Zhejiang University, Hangzhou 310003, China; Department of Medicine, University Arkansas for Medical Sciences, Little Rock, Arkansas 72205, USA |
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Abstract To construct and identify further a recombinant of Adeno-associated virus and interferon-gamma for gene therapy, the full-length IFN-γcDNA containing signal peptide was amplified by PCR, and then cloned into the pUC18. After screening, the fragment from the positive clone was then subcloned into pwpl9. After the correct recombinant was identified by digestion with SacI and BamHI, it was transfected into lymphocyte cell line H9 mediated by calcium phosphate, and the expression of IFN-γ was detected by RT-PCR and ELISA. The result showed that the IFN-γ were expressed in the H9 cells transfected with pwp/IFN-γ. The so constructed recombinant plasmid pwpl9/IFN-γ containing the full-length IFN-γ gene was expressed in mammalian cells.
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Received: 06 January 2001
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