人纤溶酶原激活剂抑制物1(PAI-1)基因与复制缺陷型腺病毒载体AdCMVHSgD重组,与pJM17质粒共转染293细胞,采用不铺琼脂的方法产生重组病毒.PCR证实PAI-1基因重组进入腺病毒.进一步感染B16(F10)细胞,细胞表面洗提物和上清分别经纤维显示胶和反向纤维蛋白显示胶检测PAI-1抑制纤溶酶原激活剂(PA)的活性.
Human plasminogen activator inhibitor 1 (PAI-1) gene was recombined into a replication-defective adenovirus vector AdCMVHSgD, co-transfected with plasmid pJM17 into human embryo kidney cell line 293 cells. Recombinant virus was produced without use of agarose overlay, identified by PCR analysis and used to infect B16 (F10) cells. The inhibition of plasminogen activator (PA) activity of PAI-1 was revealed in the elute and supernatant of infected B16 cells by regular and reverse agarose-fibrin autography.
宋海云,张菁,徐贤秀.表达人PAI-1的重组腺病毒的制备及检测[J].生物化学与生物物理进展,1999,26(6):591-593
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