Gateway Cloning of APS1 Promoter Variants for Functional Expression Analysis
This study explores the Gateway cloning technique to construct various APS1 promoter variants, including APS1 and mutant versions. The analysis is aimed at understanding the functional implications of these variants on gene expression. The pDONR and pK2GW7 vectors allow for efficient recombination, and the use of different selection markers ensures successful cloning. Supplemental Figure 6 provides detailed results of the cloning process and expression analysis, showcasing the various constructs generated for functional studies.
Gateway Cloning of APS1 Promoter Variants for Functional Expression Analysis
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Presentation Transcript
attL2 Zeo(R) EM7 promoter APS1 pDONR APS1 3639 bp attB2 T35S APS1 RB attB1 attL1 p35S p35S APS1 T35S attR2 11046 bp ccdB RB Cm(R) Kan attR1 p35S LB pK2GW7 11135 bp Sm/SpR Kan LB Sm/SpR APS1 attB1 attB2 (PCR product) O1 O2 Gateway BP reaction attP1 EM7 promoter Gateway LR reaction Zeo(R) pDONR/Zeo 4291 bp attP2 ccdB Cm(R) Supplemental Figure 6
attL2 Zeo(R) EM7 promoter APS1mut pDONR APS1mut 3639 bp attB2 T35S APS1mut RB attB1 attL1 p35S p35S APS1mut T35S attR2 11046 bp ccdB RB Cm(R) Kan attR1 p35S LB pK2GW7 11135 bp Sm/SpR Kan LB Sm/SpR Gateway LR reaction Supplemental Figure 6
