vector/plasmid selection for insert

tony.wright at csb.ki.se tony.wright at csb.ki.se
Thu Jan 19 02:07:33 EST 1995


>Hello, I am interested in a cloning vector which allows direct selection for 
>DNA inserts as pCS19 developed from Christian Sengstag (Gene 124,p141-2). 
>However, in pCS19 a BamHI site is used for subcloning. Because we want to 
>insert Klenow treated PCR fragments we are interested in a corresponding 
>vector but containing a blunt cut restriction site for subcloning (has not to 
>be a yeast shuttle vector for our purpose). Is such a vector available and 
>who may provide us with this. Could you send your answer also directly to my 
>e-mail box (hilt at po.uni-stuttgart.de) Thank you. wohi 


There is a yeast shuttle vector (pWH5) that may cater for your needs that
was published by

Wright APH, Maundrell K, Heyer W-D, Beach D, Nurse P (1986) Vectors for the
construction of gene banks and the integration of cloned genes in S.pombe
and S.cerevisiae. Plasmid, 15:156-158.

pWH5 contains a tet resistence gene driven by the lambda Pr promoter and it
also contains a version of the cI repressor gene containing cloning sites.
Cloning inserts in these sites inactivates the cI protein, thus allowing
expression of tet resistence. One of the cloning sites is SmaI.

If you would like to use this plasmid let me know and I will send it to you.

Best wishes

Anthony Wright, D. Phil.
Associate Professor
Dept of Biosciences at Novum
Karolinska Institute
Novum
S-141 57 Huddinge
Sweden
+46 8 6089159: +46 8 7745538 (fax)
tony.wright at cbt.ki.se




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