PURPOSE: To obtain miraculin having taste-modifying action and useful as a new-type sweetener, etc., by synthesizing a DNA sequence coding the amino acid sequence of miraculin, integrating the DNA sequence into a manifestation vector, transforming a cell and culturing the transformed cell.
CONSTITUTION: A miraculin gene having unknown nucleic acid sequence is designed based on the amino acid sequence of miraculin. A DNA sequence coding a protein containing the whole or a part of the amino acid sequence of miraculin is divided into DNA fragments each containing 30-60 bases. The fragments are chemically synthesized, subcloned and successively cloned to manifestation vector having E.coli tryptophan promoter. The cell of E.coli, etc., is transformed with the obtained manifestation vector containing the chemical synthesis gene of miraculin, the transformed cell is cultured in a medium, the cultivation product is centrifuged and the collected cells are subjected to bacteriolysis by ultrasonic treatment, etc. Finally, the protein is recovered from the product to obtain the objective miraculin.
URAGAMI KENICHI
SAKAMOTO KAZUCHIKA
MIKI KEIZABURO
KURIHARA YOSHIE
KURIHARA YOSHIE