The invention belongs to the technical field of
plant genetic engineering and specifically relates to a method for screening a rice
tissue specific expression type cis-acting element and a flanking sequence thereof. According to the information in a rice whole-growth-period expression profile
chip database, all
tissue specific expression genes are classified according to expression patterns. According to the information in a
plant cis-acting element
database, cis-acting elements in the
promoter regions of different rice
tissue specific expression genes are scanned and counted. After that, cis-acting elements in the
promoter regions of green tissue specific expression genes, which occur at a
high frequency, are selected as candidate elements related to green tissue specific expression. Meanwhile, the flanking sequence of one candidate element GEAT is subjected to bioinformatic prediction. The combination of the above element and the flanking sequence thereof is integrated with -46 Minimal 35S
promoter, so that the
gene expression of a GUS
gene is driven for function identification. The GUS analysis shows that, the GUS
gene is associated with the green tissue specific expression. Through the site-
directed mutagenesis experiment of the flanking sequence, a basic group having a critical role in maintaining the function of GEAT is found.