>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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