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yeast cloning vectors

GIETZ at bldghsc.lan1.umanitoba.ca GIETZ at bldghsc.lan1.umanitoba.ca
Fri Apr 23 14:17:00 EST 1993


Hi Fatima and other Ynetters:
   What we need is someone with a bit of time and a
computer to piece all these vectors together and 
submit this to the vector data bank. Will they even be accepted
these types of sequences in the database. The problem is
 this will probably be a thankless job (much like making vectors).
     On your second question about YEplac vectors copy number,
 I should be able to tell you as I built them but I have not done the
definative experiment to determine copy number (Southern
blot).  I did do a non selective segregation loss analysis. 
The results are listed in Gietz and Sugino 1988 Gene 74:pp527-534.
and they are lost at comparable levels to YEp24 and YEp213.
This runs at about 1% per generation for all the Yeps I tested!
In fact YEplac112 had a very low level of loss (0.5%).
This suggests similar copy number but like I mentioned I didn't do
the Southern.  If someone has please enlighten me! 
 Thanks for mentioning my vectors on the net!
They suffered from a lack of exposure for years but now it seems
that more people are realizing the advantages to having a yeast
ecoli shuttle vector with a completely unique pUC19 multicloning site.
Anyone needing these vectors just send me a letter or call at the address
below.  I do have the composite sequences for all of the YXplac vectors which
is available on request!

Dan Gietz Ph.D.
Assistant Professor
Department of Human Genetics
University of Manitoba
770 Bannatyne Ave, Rm 250
Winnipeg, Man, Canada
R3E 0W3
Tel.:(204)788-6458
Fax.:(204)786-8712
GIETZ at BLDGHSC.LAN1.UMANITOBA.CA
_________________________________________
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REPLY FROM: Gietz, Dr. Dan
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From: FATIMA at AIMP.UNA.AC.AT
Subject: yeast cloning vectors
Date: 23 Apr 93 15:48:00 GMT
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Dear Yeasters,

I agree with R. Gietz's note that it would be a good idea having the
sequences of common yeast vectors available; and I would be happy 
finding YEp13 on the list... This plasmid is great for multicopy
supressor screens but mapping the inserts is a pain (according to every-
one around me who tried). Moreover, one cannot simply cut out the inserts.
I am just considering making a new multicopy library in some vector
easier to handle, e.g. in one of the YEplac plasmids. Does anyone know
what is the copy number of YEplacs or some other polylinker- containing
2 micron derivatives in comparison to YEp13? 

Thanks in advance for any data or suggestions!

                         Fatima C. (fatima at aimp.una.ac.at)




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