Proceedings of the XLVII Italian Society of Agricultural Genetics - SIGA Annual Congress

Verona, Italy - 24/27 September, 2003

ISBN 88-900622-4-X

Poster Abstract - 1.31

ISOLATION AND CHARACTERIZATION OF CELL CYCLE GENES IN NICOTIANA TABACUM

M. ATALLAH, M. L. SAVO SARDARO, E. PORCEDDU

Dipartimento di Agrobiologia e Agrochimica, Università degli Studi della Tuscia

cell cycle, cDNA-AFLP, Cdc20, d123

Cell division is a basic characteristic of all living organisms.

A cDNA-AFLP approach was used to study gene expression in synchronized cell culture of Nicotiana tabacum TBY2. 20 fragments showing cell cycle regulated expression were excised from the gel, cloned and sequenced. Database comparison of these sequences identified high similarity to genes directly involved in cell cycle regulation.

The Cdc20 (the homolog of Fizzy in yeast and vertebrates) is a WD proteins involved in the M Phase, a key factors for function and regulation of the APC/C complex. APC-Cdc20 activity is present during mitosis and initiates the metaphase-anaphase transition. Cdc20 is regulated by the spindle assembly checkpoint monitoring the attachment of each chromosome to the mitotic spindle.

D123 is a novel 44-kDa gene product, which has been proposed as necessary for S-phase entry of the cell cycle, A point mutation of D123 resulted in a temperature-sensitive arrest in G1-phase in mammalian system. The protein has several potential phosphorylation sites, is highly hydrophilic, and may be highly structured in alpha-helix.

For molecular characterization of these two gene we have made several constructs for gene overexpression, gene silencing and GFP fusion. This constructs have been introduced into Nicotiana tabacum plant and inTBY2 cell suspension.