lambda gt10 vs gt11

Morris F. Manolson mm6y+ at andrew.cmu.edu
Sat Mar 20 12:13:43 EST 1993


> 
>  I'm planning to construct a cDNA library using lambda phage.  Although I
>don't have a good antibody, I'm considering using lambda gt11 instead of 
>gt10 since I may obtain good antibodies in the future.  I understand
>lambda gt11 can be screened with nucleic acids as readily as gt10.  So I
>don't see any advantages using lambda gt10.  All thoughts on this strategy
>will be appreciated.  Thanks.

The only disadvantage I know of creating a lambda gt11 library is that 
I seem to recall that in the original Young and Davis paper (which
I can't seem to find at the moment) they said that the lacZ promoter 
was slightly "leaky", ie:  you get low levels of expression of your fusion
protein without adding IPTG.  If your "protein of interest" is highly
toxic (ie: a channel),  it may never be expressed in a gt11 library
because the low levels of fusion protein expression may kill any host.
As lambda gt10 does not express the protein, there will be no selection
against toxic fusion proteins.  Thus Young and Davis suggest using
lambda gt10 if one is just going to screen by nucleic acid hybridization.

I have had three friends use the LamdaZAP vectors with good success.
This vector saves you the trouble of having to do a phage prep
when you have to take your insert out of the lambda phage to clone
into a plasmid.  Saves at the least a couple of days work.

Before you start making your own library, have you seen all the
libraries that Clontech has for sale?  They are about $400 but
they can save you a couple of weeks of work.  I purchased a 
yeast genomic gt11 library from them last year and got my gene
in a week.  Good luck in your screen!   Morrie
  

******************************************************************************
Morris F. Manolson                     Tel:  416-813-6662  (office)
Division of Cell Biology                     416-813-5729  (lab)
Hospital for Sick Children                   416-813-5028  (FAX)
88 Elm St., McMaster building        email:  Morrie at resunix.ri.sickkids.on.ca
Toronto, Ontario, Canada
M5G 1X8



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