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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:


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.

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:

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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