First published online March 8, 2002; 10.1105/tpc.010468
The Plant Cell, Vol. 14, 713-726,
March 2002, Copyright © 2002,
American Society of Plant Biologists
Use of the Transposon Ac as a Gene-Searching Engine in the Maize Genome
Matthew Cowperthwaitea,
Wonkeun Parka,
Zhennan Xua,b,
Xianghe Yan1,a,
Steven C. Mauraisa and
Hugo K. Dooner2,a,b
a Waksman Institute, Rutgers University, Piscataway, New Jersey 08855
b Department of Plant Sciences, Rutgers University, New Brunswick, New Jersey 08901
2 To whom correspondence should be addressed. E-mail dooner{at}waksman.rutgers.edu; fax 732-445-5735
We show here that, although genes constitute only a small percentage of the maize genome, it is possible to identify them phenotypically as Ac receptor sites. Simple and efficient Ac transposition assays based on the well-studied endosperm markers bz and wx were used to generate a collection of >1300 independent Ac transposants. The majority of transposed Ac elements are linked to either the bz or the wx donor loci on chromosome 9. A few of the insertions produce obvious visible phenotypes, but most of them do not, suggesting that these populations will be more useful for reverse genetics than for forward transposon mutagenesis. An inverse polymerase chain reaction method was adapted for the isolation of DNA adjacent to the transposed Ac elements (tac sites). Most Ac insertions were into unique DNA. By sequencing tac sites and comparing the sequences to existing databases, insertions were identified in a number of putative maize genes. The expression of most of these genes was confirmed by RNA gel blot analysis. We report here the isolation and characterization of the first 46 tac sites from the two insertion libraries.
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