<mxfile host="app.diagrams.net"><diagram name="roadmap"><mxGraphModel>
  <root>
    <mxCell id="0" />
    <mxCell id="1" parent="0" />
    <mxCell id="node1" value="研究问题：如何在大肠杆菌中功能表达未结合配体的血清视黄醇结合蛋白(RBP)并研究其折叠稳定性" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#DAE8FC;strokeColor=#6C8EBF;fontSize=12;verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="450" y="40" width="340" height="64" as="geometry" />
    </mxCell>
    <mxCell id="node2" value="策略：OmpA信号肽介导周质分泌，共表达PDI/PPI辅助折叠，添加N-乙酰-L-半胱氨酸" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#D5E8D4;strokeColor=#82B366;fontSize=12;verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="245" y="162" width="340" height="64" as="geometry" />
    </mxCell>
    <mxCell id="node3" value="关键改造：融合OmpA信号肽和His6标签，pASK61载体共表达PDI/PPI，IPTG诱导" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#FFE6CC;strokeColor=#D79B00;fontSize=12;verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="40" y="284" width="340" height="64" as="geometry" />
    </mxCell>
    <mxCell id="node4" value="结果：纯化获得apo-RBP，产量0.15mg/L，Kd'=0.21μM，荧光淬灭77%" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#D5E8D4;strokeColor=#82B366;fontSize=12;verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="450" y="284" width="340" height="64" as="geometry" />
    </mxCell>
    <mxCell id="node5" value="结果：Gdn·HCl展开显示单转变，apo中点2.0M，holo中点2.4M" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#D5E8D4;strokeColor=#82B366;fontSize=12;verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="450" y="406" width="340" height="64" as="geometry" />
    </mxCell>
    <mxCell id="node6" value="结果：apo ΔG°=31.4 kJ/mol，holo ΔG°=41.3 kJ/mol，配体显著增强稳定性" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#D5E8D4;strokeColor=#82B366;fontSize=12;verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="655" y="162" width="340" height="64" as="geometry" />
    </mxCell>
    <mxCell id="node7" value="结论：周质分泌+折叠辅助因子可在E. coli获得功能正常的apo-RBP，适用于其他脂笼蛋白" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#D9EAD3;strokeColor=#82B366;fontSize=12;verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="860" y="284" width="340" height="64" as="geometry" />
    </mxCell>
    <mxCell id="edge1" style="edgeStyle=orthogonalEdgeStyle;rounded=1;orthogonalLoop=1;jettySize=auto;html=1;endArrow=block;endFill=1;strokeWidth=1.6;exitX=0.5;exitY=1;exitDx=0;exitDy=0;entryX=0.5;entryY=0;entryDx=0;entryDy=0;" edge="1" parent="1" source="node1" target="node2">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="edge2" style="edgeStyle=orthogonalEdgeStyle;rounded=1;orthogonalLoop=1;jettySize=auto;html=1;endArrow=block;endFill=1;strokeWidth=1.6;exitX=0.5;exitY=1;exitDx=0;exitDy=0;entryX=0.5;entryY=0;entryDx=0;entryDy=0;" edge="1" parent="1" source="node2" target="node3">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="edge3" style="edgeStyle=orthogonalEdgeStyle;rounded=1;orthogonalLoop=1;jettySize=auto;html=1;endArrow=block;endFill=1;strokeWidth=1.6;exitX=0.5;exitY=1;exitDx=0;exitDy=0;entryX=0.5;entryY=0;entryDx=0;entryDy=0;" edge="1" parent="1" source="node3" target="node5">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="edge4" style="edgeStyle=orthogonalEdgeStyle;rounded=1;orthogonalLoop=1;jettySize=auto;html=1;endArrow=block;endFill=1;strokeWidth=1.6;exitX=0.5;exitY=1;exitDx=0;exitDy=0;entryX=0.5;entryY=0;entryDx=0;entryDy=0;" edge="1" parent="1" source="node2" target="node4">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="edge5" style="edgeStyle=orthogonalEdgeStyle;rounded=1;orthogonalLoop=1;jettySize=auto;html=1;endArrow=block;endFill=1;strokeWidth=1.6;exitX=0.5;exitY=1;exitDx=0;exitDy=0;entryX=0.5;entryY=0;entryDx=0;entryDy=0;" edge="1" parent="1" source="node1" target="node6">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="edge6" style="edgeStyle=orthogonalEdgeStyle;rounded=1;orthogonalLoop=1;jettySize=auto;html=1;endArrow=block;endFill=1;strokeWidth=1.6;exitX=1;exitY=0.5;exitDx=0;exitDy=0;entryX=0;entryY=0.5;entryDx=0;entryDy=0;" edge="1" parent="1" source="node4" target="node7">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="edge7" style="edgeStyle=orthogonalEdgeStyle;rounded=1;orthogonalLoop=1;jettySize=auto;html=1;endArrow=block;endFill=1;strokeWidth=1.6;exitX=0;exitY=0.5;exitDx=0;exitDy=0;entryX=0;entryY=0.5;entryDx=0;entryDy=0;dashed=1;dashed=1;dashed=1;dashed=1;dashed=1;" edge="1" parent="1" source="node5" target="node7">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="edge8" style="edgeStyle=orthogonalEdgeStyle;rounded=1;orthogonalLoop=1;jettySize=auto;html=1;endArrow=block;endFill=1;strokeWidth=1.6;exitX=0.5;exitY=1;exitDx=0;exitDy=0;entryX=0.5;entryY=0;entryDx=0;entryDy=0;" edge="1" parent="1" source="node6" target="node7">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
  </root>
</mxGraphModel></diagram></mxfile>