Statement of Work.pdf

STATEMENT OF WORK

Yeast Two-Hybrid-Based Screening Service General Information

1.0 Scope of Work:

The service provider will identify novel protein-protein interactions of candidate proteins. Candidate genes will be provided by the Crop Production and Pest Control Research Unit at Purdue University in West Lafayette, Indiana. One of the major goals of this ARS project is to identify host proteins targeted by pathogen-secreted effectors and, using this information, subsequently introduce new-to-nature disease resistance traits in cereal grains.

2.0 Background:

One of the major objectives of the Crop Production and Pest Control Research Unit is to investigate the molecular and genetic basis of disease resistance in cereal grains to fungal pathogens. To address this objective, the CPPC research unit would like to identify novel protein-protein interactions for several candidate proteins. Specifically, the CPPC would like to identify maize proteins targeted by fungal effectors as well as identify host proteins that interact with a disease resistance protein from barley. These data will provide us with future avenues of research and will help us unravel disease resistance signaling in cereal grains.

CONTRACTOR REQUIREMENTS

3.0 Technical Objectives and Tasks:

Features, Components, Accessories, Modules, Software and/or Services to be included:

• The service provider should screen a yeast two-hybrid platform to discover novel protein interactions.

• The vendor will generate a highly complex cDNA library generated from barley variety (CI16151) and screen this library to identify novel protein partners with a barley disease resistance protein

• The service provider will screen cDNA libraries generated from maize using a yeast two-hybrid platform to identify protein-protein interactions with two fungal effectors.

• The vendor must be able to identify weak or transient protein-protein interactions.

• The vendor should provide an exhaustive list of identified protein partners, the experimental interaction domain on each protein, and a confidence score to the Crop Production and Pest Control Research Unit.

• The vendor must perform additional screens if the initial results provide less than 20 positive clones

• Delivery schedule: three-to-six months from the time of sample submission

4.0 Deliverables / Schedule:

Key Deliverables

Item No. Deliverable Objective Due 1 Generate a highly complex cDNA library generated from barley variety (CI16151) and screen this cDNA library using a yeast two-hybrid platform. The vendor should provide an exhaustive list of identified protein partners, the experimental interaction domain on each protein, and a statistical confidence score. Vendor will perform additional screens if the initial results provide less than 20 positive clones.

The vendor must be able to identify weak or transient protein-protein interactions.

Identify barley protein partners that associate with a barley disease resistance protein

3-to-6 months from the time of sample submission

2 Screen a cDNA library generated from maize using a yeast two-hybrid platform.

The vendor should provide an exhaustive list of identified protein partners, the experimental interaction domain on each protein, and a statistical confidence score.

Vendor will perform additional screens if the initial results provide less than 20 positive clones. The vendor must be able to identify weak or transient protein-protein interactions.

Identify protein partners that interact with two fungal effector proteins

3-to-6 months from the time of sample submission