Soybean cDNA library Gm-c1074


Clones from this library are available from
    Biogenetic Services
    801 32nd Ave.
    Brookings, SD 57006 USA
    phone: 800 423 4163
    email: info@biogeneticservices.com

To insure that credit is given to the laboratory and scientists donating this valuable resource to the scientific community, the information provided with the library must be maintained and included with any transfer of biological materials or data derived from the library.

Source of Library (Donor):
Dr. Lila O. Vodkin       Phone:   217 244 6147       e-mail:   l-vodkin@uiuc.edu
University of Illinois at Champaign-Urbana              
Department of Crop Science        
384 Edward R. Madigan Laboratory        
1201 W. Gregory Drive        
Urbana, IL 61801        

Short Description of Library:
The cDNA library was constructed from mRNA isolated from unifoliate leaves from 9-11 day old seedlings that were induced for HR (hypersensitive response) by vacuum infiltrating plant tissue with Pseudomonas syringae pv.glycinea carrying the avrB gene (Genetics 141:1597-1604). Plant tissue (expanded unifoliate leaves) was collected at 2, 4, 8, 12, 24, 36, and 53 hrs after inoculation and their mRNA pooled equally for cDNA construction. The library was prepared using the Stratagene pBluescript II SK (+) library construction kit. Complementary DNA was synthesized from mRNA using a primer consisting of a poly(dT) sequence with an Xho I restriction site. Eco RI adaptors were ligated to the blunt-ended cDNA fragments followed by Xho I digestion. The cDNA insert is protected from XhoI digestion via methylation during first strand cDNA synthesis. The cDNA fragments were directionally cloned into the Eco RI-Xho I restriction site of the pBluescript vector. The ligated cDNA fragments were transformed into E. coli ElectroMax DH10B host cells. Plant care, inoculations, and library construction were performed by Steve Clough (Lila Vodkin lab, University of Illinois).

Transformation efficiency: 1010
Percent white colonies: 50%
Average insert size of white colonies: 1100 bp

Genotype/Cultivar:
Williams 82

Organ/Tissue:
9-11 day old seedlings that were induced for HR response by Pseudomonas syringae pv. glycinea carrying the avrB gene

Vector Information:
The cDNA library was prepared using the pBluescript II SK (+) vector (Stratagene) to generate a plasmid library. The vector provides ampicillin resistance and allows blue-white color selection for recombinant phagemids. The cloned cDNA fragment can be amplified by the polymerase chain reaction using one of the following vector primer pair combinations: T3 and T7 or M13 universal and M13 reverse.


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