VECTOR FOR ANALYSIS OF PROMOTER ACTIVITY IN HIGHER PLANT AND APPLICATION METHOD THEREOF

摘要

The development of knock-in method by homologous recombination for qualitative and quantitative analyses of a promoter in a higher plant has been strongly demanded. The invention is directed to a vector for analysis of a promoter activity in a higher plant constructed by including the following elements between T-DNA-derived RB and LB in the order of the following description from RB: (1) a first negative selectable marker gene which is allowed to exist in an expressible manner; (2) a first homologous recombination region consisting of a nucleotide sequence of 500 to 6000 bp containing the 3' end of a target promoter in a host genome; (3) areporter gene in which its initiation codon is located at a desired distance from the 3' end of the promoter; (4) a positive selectable marker gene which is allowed to exist in an expressible manner; (5) a second homologous recombination region consisting of a nucleotide sequence of 500 to 6000 bp followed by the first homologous recombination region in the host genome; and (6) a second negative selectable marker gene which may be the same as the first negative selectable marker gene allowed to exist in an expressible manner. By searching a promoter highly expressed in a tissue which is easily extracted or the like using this vector and ligating a useful protein gene or a gene related to the production of a useful substance thereto, mass production of a useful protein and the like can be realized.