<mxfile host="app.diagrams.net"><diagram name="roadmap"><mxGraphModel dx="1208" dy="795" grid="1" gridSize="10" guides="1" tooltips="1" connect="1" arrows="1" fold="1" page="1" pageScale="1" pageWidth="1200" pageHeight="800" math="0" shadow="0">
  <root>
    <mxCell id="0" />
    <mxCell id="1" parent="0" />
    <mxCell id="node1" value="核心问题：lpldh表达调控机制及高效LPLDH重组菌" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#dae8fc;strokeColor=#6c8ebf;fontSize=13;fontStyle=1verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="406" y="40" width="334" 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;strokeColor=#6c8ebf;" edge="1" parent="1" source="node1" target="node2">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="node2" value="策略：分析lpldh上下游序列，发现TetR家族lplR基因" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#d5e8d4;strokeColor=#82b366;fontSize=12verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="402" y="162" width="340" height="64" 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;strokeColor=#82b366;" edge="1" parent="1" source="node2" target="node3">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="node3" value="关键改造：构建ΔlplR菌株及多拷贝lplR质粒pKLplR" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#dae8fc;strokeColor=#6c8ebf;fontSize=12verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="198" y="284" width="340" height="64" 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;strokeColor=#6c8ebf;" edge="1" parent="1" source="node3" target="node4">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="node4" value="结果验证：EMSA证明LplR结合6-bp反向重复序列" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#fff2cc;strokeColor=#d6b656;fontSize=12verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="40" y="406" width="320" height="64" 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=1;entryDx=0;entryDy=0;strokeColor=#d6b656;" edge="1" parent="1" source="node4" target="node5">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="node5" value="关键结论：敲除lplR使lpldh表达量提高22.8倍" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#f8cecc;strokeColor=#b85450;fontSize=12verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="792" y="406" width="313" height="64" 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;strokeColor=#b85450;" edge="1" parent="1" source="node5" target="node6">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="node6" value="最终应用：ΔlplR(pKLPLDH)菌株用于D-PL立体转化" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#d5e8d4;strokeColor=#82b366;fontSize=12verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="608" y="580" width="340" height="64" 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=0;exitY=0.5;exitDx=0;exitDy=0;entryX=0;entryY=0.5;entryDx=0;entryDy=0;dashed=1;dashed=1;dashed=1;strokeColor=#82b366;dashed=1;" edge="1" parent="1" source="node5" target="node7">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="node7" value="结果指标：LPLDH活性16.3 mU/mg，极显著优于野生型" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#ffe6cc;strokeColor=#d79b00;fontSize=12verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="608" y="284" width="340" height="64" 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=0.5;exitY=1;exitDx=0;exitDy=0;entryX=0.5;entryY=0;entryDx=0;entryDy=0;strokeColor=#d79b00;dashed=1;" edge="1" parent="1" source="node7" target="node5">
      <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;strokeColor=#b85450;" edge="1" parent="1" source="node2" target="node7">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="node8" value="机制阐明：LplR结合IR序列抑制操纵子转录" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#e1d5e7;strokeColor=#9673a6;fontSize=12verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="430" y="406" width="292" height="64" as="geometry" />
    </mxCell>
    <mxCell id="edge9" 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;strokeColor=#9673a6;" edge="1" parent="1" source="node7" target="node8">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="edge10" 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;strokeColor=#9673a6;" edge="1" parent="1" source="node3" target="node8">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="node9" value="应用延伸：E. coli共表达KPR/GDH实现D-PA高效合成" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#f8cecc;strokeColor=#b85450;fontSize=12verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="198" y="580" width="340" height="64" as="geometry" />
    </mxCell>
    <mxCell id="edge11" 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;strokeColor=#b85450;" edge="1" parent="1" source="node8" target="node9">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
    <mxCell id="node10" value="结论：LplR为负调控因子，敲除可实现不依赖诱导剂的LPLDH高表达" style="rounded=1;whiteSpace=wrap;html=1;fillColor=#d5e8d4;strokeColor=#82b366;fontSize=13;fontStyle=1verticalAlign=middle;align=center;" vertex="1" parent="1">
      <mxGeometry x="402" y="702" width="340" height="64" as="geometry" />
    </mxCell>
    <mxCell id="edge12" 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;strokeColor=#82b366;" edge="1" parent="1" source="node9" target="node10">
      <mxGeometry relative="1" as="geometry" />
    </mxCell>
  </root>
</mxGraphModel></diagram></mxfile>