New ABRC DNA stock

Keith R Davis krdavis at magnus.acs.ohio-state.edu
Mon Nov 14 11:28:02 EST 1994


The ABRC is pleased to announce the availability of a new library stock that is
suitable for use with the GAL4 interaction tap system for analyzing
protein-protein interactions.  Many thanks to John Walker (University of
Missouri) for making this library available.

Stock# CD4-10
Expression cDNA library

This kit includes the yeast strain, vectors, maps and cDNA library in the pACT
vector (activation domain) which will allow you to screen for interacting
proteins in a yeast two-hybrid system.  These vectors were developed by Steve
Elledge at Baylor College of Medicine.  For more details please refer to:

Durfee et al. (1993) The retinoblastoma protein associates with the protein
phosphatase type 1 catalytic subunit.  Genes & Development 7:555-569.

The kit includes:

Yeast strains

1    Y190, slant
1    Y190 (pAS1 + pCL1) GAL4 control, slant
1    Y190 (pSE1111 + pSE1112) SNF1/SNF4 control, slant

plasmid DNA
1    pAS1, dried pellet
1    pAS1-CYH2, dried pellet

cDNA library
1    cDNA library in vector pACT with host BNN132, 7% DMSO stock


The lambda ACT library was made with random-primed mRNA isolated from mature
Arabidopsis leaves and roots.  The blunted cDNA was ligated to adaptors:

GGCCTTCGTGGCC
CCGGAAGCACCGGAGC

and size selected for inserts > 300 bp.  The adapted cDNA was inserted into the
T-filled XhoI site of lACT.  1.9 x 106 primary transformants were obtained, at
least 70% are recombinant.  BNN132  was infected with 300 ul of 7
x108/ml amplified stock.  After infections the library/host was transfered
to 1 L of TB broth.  30 minutes after innoculation  50 ml of the culture was
plated on LB glucose amp plates for a total 2600 colonies (5.2 107) cfu.
The culture was grown overnight and used to prepare DMSO stocks for shipping.
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All DNA stocks can be ordered from the ABRC DNA Stock Center by e-mail
(dna at genesys.cps.msu.edu), by FAX (614-292-5379) or by regular mail sent to:

ABRC
Biotechnology Center
1060 Carmack Road
Columbus, OH  43210
Attn: Doreen Ware

Any comments or suggestions concerning the DNA Stock Center can be directed to
Keith Davis (krdavis at magnus.acs.ohio-state.edu).



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