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Ura3-52 deletion in Clontech strains

cgdn at djembe.biophysics.rochester.edu cgdn at djembe.biophysics.rochester.edu
Wed Apr 2 09:43:20 EST 1997


On 25 Mar 1997 14:22:50 GMT, lichten at helix.nih.gov (Michael Lichten)
wrote:

>In article <espinoza.859230092 at cgl.ucsf.edu>, espinoza at cgl.ucsf.edu
>(Hernan Espinoza) wrote:
>
>> Albert Spielmann <Albert.spielmann at bota.unine.ch> writes:
>> 
>> >I'm working with the two-hybrid system from Clontech.
>> >The yeast strains are CG1945 and Y190. Both have a ura3-52 deletion.
>> >Normally they can't grow without uracil. But as said in the supplier's
>> >protocol, it must grow!! Why?
>> 
>>         Your question is a bit ambiguous, under what conditions must the
>> strains grow?  I'm not familiar with the Clonetech system, but I would
>> wager it's to select for a URA3 marker on a plasmid...
>> 
>>         -Hernan
>
>Let's clear up a misconception right now.  ura3-52 is NOT a deletion; it's
>a Ty insertion in the URA3 coding region.
>
>-- 
>Michael Lichten
>lichten at helix.nih.gov

The yeast strain Y190 DOES carry the ura3-52 marker at the ura3 locus.
However this strain is URA+ because it contains a WT URA3 gene which
was used as a marker to insert the GAL->lacZ sequence. This sequence
is composed of the GAL promoter linked to the E. coli lacZ coding
sequence. It is used as a color stain to determine when the GAL
promoter has been activated by a two hybrid interaction.



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