removing NOS:NPTII:NOS from T-DNA region

Chung-Jui Tsai chtsai at MTU.EDU
Thu Nov 5 14:38:46 EST 1998

You can try call Clontech up to request the complete DNA sequence of
pBI121, although they no longer carry this series of vectors.  Or you can
use PmeI (a blunt-end cutter) and HindIII to remove the NPT-II cassette.
PmeI is located near (~10 bp) the 5'-end of the NOS promoter, or ~35 bp
from the beginning of the right border sequence.  The T-DNA sequences of
or pBI101 series ARE available in the genbank database but not their entire
vector sequences.

Hope this helps.

Chung-Jui Tsai
Michigan Tech University

At 01:46 PM 11/4/1998 -0800, you wrote:
>I was wondering if any one has tried to remove the selectable marker
>NOS:NPTII:NOS casette from pBI121.1 or any of its parent vectors and could
>offer advice.  I would like to place a new selectable marker in its place
>and I can not find a detailed restriction map of the T-DNA region.  I
>tried to construct a map using the genes and ori's present in the gene
>bank for the T-DNA region and pBIN19.  However, I did not feel confident
>in using this aproach for experimentation because the gene sequences I
>found would not line up right when using megaline to match gene to vector.
>Please send any knowledge of restriction maps, references, gene sequences,
>or methods on how to remove NPTII from the T-DNA region
>Thanks you very much
>Alan Smith  

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