Question about pCLEAN plasmid

[Talon]

I've been looking for binary vectors for the transformation of Agrobacterium and the eventual transformation of a plant. I came across a couple binary vectors, but I found pCLEAN to have the most suitable vector choices. When I was looking at the different vectors they contain on their website, I noticed some choices would give you T-DNA on both plasmids. What is the purpose of having two T-DNAs?

 

Also when I was trying to determine which vector of pCLEANto use, I came across this image in a paper evaluating pCLEAN vectors

The pictures of G130, G156, and G184 GFP expression don't make sense to me, as how did the GFP gene get into the plant if it was located outside the T-DNA borders?

 

 

For quick reference (it is in the link provided below) here is the legend for the picture above

Here is the pCLEAN information http://www.jic.ac.uk...ctor_system.pdf.

Edited October 30, 2010 by Talon

[CharonY]

I assume it should be noted in a paper somewhere describing those plasmids. However, it is known that sometimes the backbone gets transfected. Judging from the pictures I assume that it was conducted to assess the rate of backbone transfer.

Also, the additional T-DNA could be exploited for the delivery of additional sequences.

[Talon]

I know that the backbone gets transfered now, but how does this happen? As for the multiple T-DNA strans, it was so you could create a marker free plant. Plants transformed by two T-DNAs can be selected out to express only one of the T-DNAs. Pictures were obtained from http://www.plantphysiol.org/cgi/content/full/145/4/1211#BIB15 .

Edited October 31, 2010 by Talon