Useful All Purpose Yeast:coli Vectors

odile at FRCGM51.EARN odile at FRCGM51.EARN
Sun May 9 07:22:35 EST 1993


 
 
Hello  Yeast netters,
 
Daniel wrote :
>.....
>.......
>I removed the EcoRI and KpnI sites from the LEU2 gene
>as well as the HindIII, XbaI and PstI sites from TRP1 ARS1 frag
   for each selectable marker.
>
>SEE Gietz and Sugino (1988) Gene 74:527-534
>
>  Selectable Marker    TRP1       URA3        LEU2
>
>ARS1 CEN4 plasmid    YCplac22    YCplac33    YCplac111
>2 micron plasmid     YEplac112   YEplac195   YEplac181
>Integrating plasmid  YIplac204   YIplac211   YIplac128
 
........
>I have also compiled DNA sequence file for each vector made by piecing
> together all the fragments. They too are available for research purposes.
 
 
Bravo for your shuttle vectors !! We also constructed a shuttle vector
family ( the pFL vectors) looking like yours but with in add,
7 other plasmids :
- 3 multicopy vectors bearing the STB gene from 2 microns that conferres
a higher stability than the multicopy plasmids bearing only the 2 micron
origin,
- 4 plasmids bearing also the origin of replication of the
filamentous bacteriophage f1 into a centromeric (TRP) and a multicopy
vector (URA) yielding single stranded DNA in E.coli.
 
Ref : Bonneaud et al, 1991 "A family of low and high replicative,
integrative and single-stranded S.cerevisiae/ E.coil shuttle vectors",
Yeast, vol 7, 609-615
 
But Daniel, you were more courageous than us for the LEU2 marker,
because we did not destroyed its ecor1 and kpn1 sites .. ;-)
 
When we published our paper, I sent the sequences in Vecbase....
but as known, Vecbase does not appear alive...
so I sent the sequences at merlot.welch.jhu.edu (Dan Jacobson)
where they are accessible by gopher.
Nevertheless,  it should be a good thing if vecbase (or
another data bank) could publish a new release for vectors,
because its accessibility is easer than by gopher (it's my
opinion...;-) ). Is any one know if vector sequences
are the welcome in a easy data bank (at my knowledge, EMBL
does not accept vectors and for Gene bank, I don't know...)
 
 
Au revoir,
 
Odile Ozier-Kalogeropoulos
CGM CNRS
91190 Gif sur Yvette
FRANCE
 
E-mail : Odile at CGMVAX.CGM.CNRS-GIF.FR
 
 
 
 
 
 
 



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