摘要
目的 :探讨两种基因转移载体脂质体转染的和重组腺病毒收集并转导的SEAP基因在B16F1细胞中的表达动态。方法 :脂质体和质粒DNA复合后 ,转染 10 6 B16F1细胞 ,连续 5d收集并换细胞培养液 ,并检测其中的SEAP酶活性。将重组腺病毒以 10 0 0m .o .i、2 0 0 0m .o .i、3 0 0 0m .o .i、4 0 0 0m .o .i接种B16F1细胞 ,连续 5d收集并换细胞培养液 ,并检测其中SEAP的含量。结果 :随着时间的推移 ,脂质体转染的SEAP的表达逐渐下降 ,由第 1天的 82 1.6μUnit/ml下降到第 5天的 2 75 .2 μUnit/ml。腺病毒转导的SEAP的表达却是逐渐上升。并随接种毒量的增加而上升。且第三、四天表达达最高后 ,能保持较稳定的水平。接种 4 0 0 0m .o .i组 ,SEAP的表达由第 1天的93 .9μUnit/ml上升到第 3天 12 81.1μUnit/ml。到第 5天仍为 12 4 0 .4 μUnit/ml。结论 :脂质体和重组腺病毒作为不同的基因转移载体都能有效地介导外源基因的转移入靶细胞 ,并使外源基因在靶细胞内获得有效地表达。
Objective: To compare the expression kinetics of SEAP gene from B16F1 cells transfected by cationic lipid and transducted by recombinant adenovirus in order to explore the potential application of two gene transfer vector: cationic lipid and recombinant adenovirus.Methods:The lipid DNA complex was added to B16F1 cells of 106. The suspernatant of cell culture was collected and replaced each day for 5 days and SEAP levels were assayed with SEAP kit.The recombinant adenovirus at m.o.i 1000?m.o.i2000?m.o.i3000?m.o.i4000 was inoculated respectively to B16F1 cells of 106. Results: The expression level of SEAP decreased from 821.6 μUnit/ml on day 1 to 275.2 μUnit/ml on day 5 gradually.The expression level of SEAP elevated gradually with increasing m.o.i of inoculated adenovirus and time going. In the group inoculated with adenovirus at m.o.i 4000,SEAP level increased from 93.9 μUnit/ml on day 1 to 1281.1 μUnit/ml on day 3 and 1240.4 μUnit/ml on day 5.Conclusion:Cationic lipid and recombinant adenovirus,as two different gene transfer vectors,could deliver foreign SEAP genes into B16F1 cells in which SEAP expressed efficiently.but both expression kinetics are completely different.
出处
《江西医学院学报》
2002年第2期15-18,共4页
Acta Academiae Medicinae Jiangxi
