摘要
目的：选择适当的载体，启动Ｃａ＾２＋依赖分泌型磷脂酶Ａ２（ｓＰＬＡ２）ｃＤＮＡ的表达，为进一步研究各种因素对该酶活性的影响奠定基础。方法：首先从ｓＰＬＡ２－ｐＧＥＭ７重组体（这一重组体不能在转染的真核细胞中表达）获得人ｓＰＬＡ２ｃＤＮＡ，克隆该片断进入中间载体ＢＳＳＫ（使ｓＰＬＡ２ｃＤＮＡ两端出现ＸｂａＩ、Ｈｉｎｄ３酶切位点），选择ａｎｔｉｓｅｎｓｅ ｓＰＬＡ２－ＢＳＳＫ重组体进行扩增。
Objective:Choose the effective vector for sPLA2cDNA and study the expression of sPLA2 in the vector.Method:we prepared the sPLA2cDNA from pGEM7; lagation of aPLA2-BSSk for getting different cohesive ends of sPLA2cDNA by Xbal HindIII: purifying the sPLA2-PRC/CMV;trasfacted sPLA2-PRC/CMV with mouse macrophage;isolatied RNA from macrophage cells,Northem blot;autoradiograph.Results:the orientation lagation of insert DNA is useful and essy to get recombinstion DNA;pGEM/is a expressing vector but the aPLA2 cDNA in it couldn't be expressed .Conclusion:PRC/CMV is a useful expressing vector for aPLA2 cDNA
