Du Lab

 

New reverse genetics resources for maize:
facile production and efficient indexing using next-generation sequencing technology

 

Project Description

A sequence-indexed reverse genetics resource is considered essential to fully exploit the maize genome sequence released in 2009. Our approach to creating such a resource entails generating, sequencing, and indexing new Ac and Ds insertions in maize. Under funding from the NSF Plant Genome Program, our project is developing a general method for sequence-indexing new insertions that is rapid, accurate, and cost-effective by taking advantage of sample pooling and next-generation sequencing technologies.

 

Aim of Project

  • To sequence-index a collection of 1150 existing Ac transposants described in Cowperthwaite et al (2002) The Plant Cell 14, 713-726.
  • Create a set of 120 roughly equidistant transgenic Ds* launching platforms carrying easily scored markers, such as GFP, that will allow simple visual selection of element transposition from any region of the genome and, thus, enable researchers to generate regional gene knock-out collections;
  • Sequence-index 10,000 Ds* insertion sites from model platforms using a novel method that should be generalizable to any collection of insertions produced in a common genetic background;
  • Develop a web-searchable database of insertion site sequences cross-referenced to stocks available from the Maize Genetics Stock Center.