Award Abstract #0211547
A Set of Maize Transgenic Lines for Localized Mutagenesis Based on the Ac-Ds Transposon System Mutagenesis Based on the Ac-Ds Transposon System

NSF Org:	IOS Division of Integrative Organismal Systems
Initial Amendment Date:	August 2, 2002
Latest Amendment Date:	August 2, 2002
Award Number:	0211547
Award Instrument:	Standard Grant
Program Manager:	Jane Silverthorne IOS Division of Integrative Organismal Systems BIO Directorate for Biological Sciences
Start Date:	September 15, 2002
Expires:	September 30, 2004 (Estimated)
Awarded Amount to Date:	$140612
Investigator(s):	Hugo Dooner dooner@waksman.rutgers.edu (Principal Investigator)
Sponsor:	Rutgers University New Brunswick 3 RUTGERS PLAZA NEW BRUNSWICK, NJ 08901 732/932-0150
NSF Program(s):	PLANT GENOME RESEARCH PROJECT
Field Application(s):
Program Reference Code(s):	BIOT, 9109
Program Element Code(s):	1329

ABSTRACT

The goals of this NSF Plant Genome project are to demonstrate that the transposon Ac (Activator) can be used as a gene identification and isolation tool, as well as a mutagen, in the complex maize genome, and to develop the necessary tools to facilitate that use. The first goal was met in the initial phase of this project where Ac was shown to insert exclusively in or close to genes and to be, therefore, an excellent gene-searching engine in the highly repetitive maize genome. A highly embryogenic bz wx inbred line was developed and transformed with Ac* and Ds* constructs that had been modified to facilitate the PCR isolation of the transposon-adjacent sequence. The main goal of the second phase of this project is to create a comprehensive set of transgenic lines that will serve as starting points for the production of future insertion libraries. This phase will occur in two well-defined stages. The first will be to demonstrate the germinal transposition of an engineered Ac* or Ds* element and the ready isolation of DNA adjacent to the transposon. The second will be to identify a method of maize transformation that will enable the production of a useful set of transposon lines for localized mutagenesis. Once the above two objectives have been met, the last stage of the project to produce a set of 124 transgenic lines with a uniquely marked element at evenly spaced locations in the genome will be initiated. In that set, most genes in the maize genome will be within 7 cM of a launching platform and will be, therefore, realistic targets in a localized transposon mutagenesis experiment. These lines will be deposited in the Maize Stock Center and will serve as starting materials for the generation of future insertion libraries by interested scientists.

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