Poster: Emerging Technologies

Abs # 883: Construction of the advanced system for insertional mutagenesis using transposon with GFP.

Presenter:	Tanimoto, Takayuki , t01h206@tmail.shinshu-u.ac.jp
Authors	Tanimoto, Takayuki  (A)   Iwata, Fumi  (A)   Nakanishi, Hiromitsu  (A)   Taguchi, Goro  (A)   Niwa, Yasuo  (B)   Kojima, Mineo  (C)   Hayashida, Nobuaki  (A)
Affiliations:	(A): Gene Research Center, Shinshu University (B): Dept. of Food & Nutritional Sciences, Grad. School of Nutrional & Environ. Sciences, Univ. of Shizuoka (C): Dept. of Applied Biology, Faculty of Textile Science & Technology, Shunshu Univ.

Elucidation of the functions of individual gene products is the major subject of post genome era. Among several methods to produce mutant library, the transposon tagging system has possibility to develop a simple, large-scale, and reproducible mutagenesis system, with the ease of analysis after mutation. In this study, we constructed a plasmid pGTII for the purpose. pGTII bear a T-DNA consisting of two regions, transposon region and base region. The transposon region has a promoterless GFP gene and a hygromycin resistance gene (Hyg^r). The base region has a suicide gene codA (negative marker) and a herbicide BASTA resistance gene. A parent plant in which GTII T-DNA is introduced is crossed with the transposed supplying line to get progenies being active in transposition. In the nest generation, only the plants that inherited excised and re inserted transposon region but not base region would be selected with codA and Hyg^r. The expression with GFP on transposon is expected to reflect both the expression level of inserted genes and the localization of gene products in the cell architecture. We need to prerare milliones of "parental generation" that bear both Ac and GTII T-DNA to make a large mutant library that could cover hypothetically all genes of Arabidopsis genome. The three selection markers, Hygr, bar for GTII and Km^r for Ac, will help the achievement of the work.