Difference between revisions of "Part:BBa K5407000"
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===Usage and Biology===
 
===Usage and Biology===
  
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pLKO.1 plasmid is a basic backbone plasmid and can be expressed in mammalian cell lines. The plasmid map can be viewed in the figure 1.The backbone size of the plasmid is 7032 bp,and the selectable markers of the plasmid is puromycin Scientists often use the plasmid tool to inhibit the expression of genes. In our study, we cloned the siRNA sequence targeted to TEAD4 into the basic part pLKO.1 plasmid. And we constructed two composite plasmid named sh-TEAD4-1 and sh-TEAD4-2.Then the TEAD4 mRNA could be degraded in the SW480 cell.
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pLKO.1 plasmid is a basic backbone plasmid and can be expressed in mammalian cell lines. The plasmid map can be viewed in the figure 1.The backbone size of the plasmid is 7032 bp,and the selectable markers of the plasmid is puromycin. Scientists often use the plasmid tool to inhibit the expression of genes. In our study, we cloned the siRNA sequence targeted to TEAD4 into the basic part pLKO.1 plasmid. And we constructed two composite plasmid named sh-TEAD4-1 and sh-TEAD4-2.After we transfected the plasmid into  SW480 cell, the TEAD4 mRNA could be degraded in the cell.
 
                                  
 
                                  
 
             https://static.igem.wiki/teams/5407/plko.png
 
             https://static.igem.wiki/teams/5407/plko.png

Revision as of 06:06, 21 September 2024

pLKO.1 puro plasmid

Usage and Biology

pLKO.1 plasmid is a basic backbone plasmid and can be expressed in mammalian cell lines. The plasmid map can be viewed in the figure 1.The backbone size of the plasmid is 7032 bp,and the selectable markers of the plasmid is puromycin. Scientists often use the plasmid tool to inhibit the expression of genes. In our study, we cloned the siRNA sequence targeted to TEAD4 into the basic part pLKO.1 plasmid. And we constructed two composite plasmid named sh-TEAD4-1 and sh-TEAD4-2.After we transfected the plasmid into SW480 cell, the TEAD4 mRNA could be degraded in the cell. 

           plko.png
                           Figure 1 pLKO.1 map


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 1645
    Illegal SpeI site found at 1182
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 1645
    Illegal SpeI site found at 1182
    Illegal NotI site found at 1931
    Illegal NotI site found at 2829
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 1645
    Illegal BglII site found at 107
    Illegal BglII site found at 148
    Illegal BglII site found at 214
    Illegal BglII site found at 3501
    Illegal BamHI site found at 925
    Illegal XhoI site found at 1444
    Illegal XhoI site found at 1634
    Illegal XhoI site found at 1938
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 1645
    Illegal SpeI site found at 1182
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 1645
    Illegal SpeI site found at 1182
    Illegal NgoMIV site found at 6296
    Illegal AgeI site found at 1669
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 426
    Illegal BsaI.rc site found at 125
    Illegal BsaI.rc site found at 1340
    Illegal BsaI.rc site found at 3519
    Illegal BsaI.rc site found at 5120
    Illegal SapI site found at 2866
    Illegal SapI site found at 4037