[Arabidopsis] influence of CaMV35S on the pattern of nearby promoter-GUS

Luoping Yan via arab-gen%40net.bio.net (by yanluoping At hotmail.com)
Tue Jan 2 21:09:25 EST 2007


>Hello researchers,
>
>I am study one gene promoter and fused it to GUS in pCAMBIA1391Z, in which 
>CaMV35S drives the selectable marker Hyg gene. Among the 10 independent T3 
>lines, which were confirmed by PCR, some show very strong GUS staining, 
>some no, and some weak staining.
>
>I knew from this paper that vectors containing 35S promoter driving 
>selectable marker gene are not good for promoter study.
>The 35S promoter used in a selectable marker gene of a plant transformation 
>vector affects the expression of the transgene. Planta. Volume 221, Number 
>4 / June, 2005
>
>But I found that quite many published papers used vectors (like 
>pCAMBIA1391, pCAMBIA1301, etc) containing 35S promoter driving selectable 
>marker.
>
>As the staining patterns of my promoter fusion lines are not consistent at 
>al, I want to use another vector pBI121 or pCGN1547. Does anyone know where 
>to get pCGN1547?
>How do you deal with those lines from 35S containing vector? CAMBIA 
>suggests doing co-transformation strategy to complete remove the influence 
>of CaMV35S, i.e. one vector without selectable marker gene in T-DNA and the 
>other contains only selectable marker  gene in T-DNA. But it will involve a 
>lot of work and screening workload would be large.
>Has anyone tried this and how about the success rate?
>
>Thanks for your input,
>
>Luoping

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