National Institute of Agrobiological Sciences

National Agriculture and Bio-oriented Research Organization

Preventec, Inc.

National Institute of Advanced Industrial Science and Technology

Nippon Paper Industries Co., Ltd.

Sanwa Kagaku Kenkyusho Co., Ltd.

Incorporated Administrative Agency National Agriculture and Food Research Organization

Fumio Takaiwa

Leqing Qu

Saori Kasahara

Kenji Sekikawa

Yoshihiro Fujiwara

Takahito Jomori

Hidenori Takagi

Hiroyasu Ebinuma

Koichi Sugita

Yuji Hayashi

Akira Tashita

Noriko Tsuji

Yuji Fujii

Yukari Kobara

Yuhya Wakasa

Yoshifumi Tada

Yasushi Kawagoe

Yasuhiko Aiki

Yasuyuki Onodera

Lijun Yang

Tomotaka Shinya

Taiji Kawakatsu

It is an object of the present invention to provide a protein that can successfully treats almost all patients with cedar pollinosis and can be ingested as a food. That is, the present invention provides a protein comprising an amino acid sequence represented by SEQ ID NO:1 or comprising an amino acid sequence having a mutation(s) in said amino acid sequence, and having an immunogenicity of a cedar pollen; a polynucleotide encoding said protein; a vector comprising said polynucleotide; a transformant comprising said polynucleotide or said vector; and intended uses of said protein, said polynucleotide, and said transformant.

Protein having immunogenicity of cedar pollen, polynucleotide encoding the protein, and use thereof

Providing a new method of treating inflammatory gastrointestinal disorders and a food product for the prevention or improvement of inflammatory gastrointestinal disorders. Prevention or treatment of inflammatory gastrointestinal disorders is possible by oral administration of recombinant IL-10 expressed in rice plant seeds.

Method of treatment of gastrointestinal disorders with IL-10

Extraction and purification of recombinant proteins rendered difficult to extract from transgenic plants by using an extraction solution containing reducing agents and surfactants or an extraction solution containing reducing agents and organic solvents.

Method for extraction and purification of recombinant proteins from transgenic plants

The present inventors identified a novel rice glutelin gene, GluD-1, which is expressed specifically in seeds. The promoter of the GluD-1 gene was confirmed to induce seed-specific gene expression, and to induce expression of downstream genes specifically in the endosperm during the early stage of seed maturation process. More specifically, the GluD-1 promoter can induce strong expression of an exogenous gene in sites including the endosperm.

Gene capable of being expressed specifically in endosperm of plant, promoter for the gene, and use of the gene and the promoter

The present invention provides a method for highly producing a recombinant protein in a plant storage organ and a GLP-1 derivative. The plant storage organ in which the recombinant protein is highly produced is obtained by transformation with the use of a vector which comprises a recombinant protein gene, a cytokinin-related gene, a drug-resistant gene and a removable DNA element, in which the cytokinin-related gene and the drug-resistant gene exist in the positions so that they can behave together with the DNA element, while the recombinant protein to be expressed in the plant storage organ exists in the position so that it would not behave together with the DNA element. The GLP-1 is produced by using the method, and a derivative having been stabilized against enzymatic digestion is further provided.

GLP-1 derivative

An objective of the present invention is to provide promoters having seed-specific promoter activity, and methods of expressing foreign proteins in seeds. The present inventors isolated the promoter of ADP-Glucose pyrophosphorylase which is expressed in rice seeds, constructed binary vectors in which the ADP-Glucose pyrophosphorylase promoter is inserted upstream of the GUS reporter gene, and transformed rice using themethod. GUS expression level was used as an index to examine the site of expression, the expression pattern during seed maturation, and the level of expression in seeds for the ADP-Glucose Pyrophosphorylase promoter. Expression was found in the embryo during early stage of maturation and in the entire seed during maturation. Expression in late stage of maturation in embryo was very high.

AGPase promoter from rice and uses thereof

An objective of the present invention is to provide promoters having seed-specific promoter activity, and methods of expressing foreign proteins in seeds. In particular, a seed specific promoter from the rice glutelin GluB-4 gene is disclosed. The present inventors isolated the promoters of a number of genes that are expressed in rice seeds, constructed binary vectors in which each promoter is inserted upstream of the GUS reporter gene, and transformed rice using themethod. The inventors then used GUS expression level as an index to examine the site of expression, the expression pattern during seed maturation, and the level of expression in seeds for each promoter. They thus discovered promoters with activity specific to a particular site in seeds, and with higher activity than constitutive promoters and known seed-specific promoters.

Seed-specific promoter from the rice glutelin GluB-4 gene and uses thereof

An objective of the present invention is to provide a promoter from the rice 10 kDa Prolaminin gene having seed-specific promoter activity, and methods of expressing foreign proteins in seeds. The present inventors isolated the promoters of a number of genes that are expressed in rice seeds, constructed binary vectors in which each promoter is inserted upstream of the GUS reporter gene, and transformed rice using themethod. The inventors then used GUS expression level as an index to examine the site of expression, the expression pattern during seed maturation, and the level of expression in seeds for each promoter. They thus discovered promoters with activity specific to a particular site in seeds, and with higher activity than constitutive promoters and known seed-specific promoters.

Seed-specific gene promoter from the rice 10 KDa prolaminin gene and uses thereof

The present inventors succeeded in developing a vector that expresses high levels of a foreign gene in plant seeds by utilizing a 5'-untranslated region of a gene encoding a seed storage protein. The inventors also succeeded in accumulating high levels of a foreign gene product in plant seeds by utilizing a seed storage protein defective mutant as a target for foreign gene transfer.

Method of accumulating foreign gene product in plant seed at high level

The present inventors presumed that rice globulins accumulating in vacuole-derived type II protein bodies comprise a vacuolar translocating signal and proceeded to identify such a signal. As a result, fusion proteins composed of a 15 amino acid residue peptide of globulin, extending from the 72nd leucine residue to the 86th serine residue, added to the C terminus of GFP were surprisingly found to be intracellularly localized to non-vacuole-derived type I protein bodies (PB-I), and not to vacuole-derived type II protein bodies (PB-II). Furthermore, based on this 15 amino acid residue sequence, the consensus sequence 'QCCXQ' (where X is an arbitrary amino acid), which is conserved in plants, was discovered. Accordingly, the present invention suggests that arbitrary peptides can be accumulated in plant endosperm tissues by adding the QCCXQ sequence thereto.

Methods for accumulating arbitrary peptides in plant protein bodies

cDNAs (RISBZ1, RISBZ4, and RISBZ5) encoding bZIP transcription factors were isolated from a cDNA library originating in rice plant seed. The cDNAs encode novel proteins and have binding activity to the GCN4 motif. Among them, RISBZ1 activated transcription mediated by the GCN4 motif by 100-fold or more. Since the expression of RISBZ1 precedes the expression of a seed storage protein gene and is expressed only in maturing seeds, it is suggested that RISBZ1 controls the expression of rice seed storage proteins. In addition, by linking the recognition sequence of the transcription factor, the GCN4 motif, in tandem and introducing it into the promoter for a gene encoding seed storage protein to facilitate its binding to the transcription factor RISBZ1, expression of a foreign gene under the control of the modified promoters is greatly enhanced.

Bzip type transcription factors regulating the expression of rice storage protein

An objective of the present invention is to provide promoters having seed-specific promoter activity, and methods of expressing foreign proteins in seeds. The present inventors isolated the promoters of a number of genes that are expressed in rice seeds, constructed binary vectors in which each promoter is inserted upstream of the GUS reporter gene, and transformed rice using themethod. In particular a seed-specific promoter from the rice glutelin GluB-1 gene is disclosed. GUS expression level was used as an index to examine the site of expression, the expression pattern during seed maturation, and the level of expression in seeds for each promoter. Promoters with activity specific to a particular site in seeds, and with higher activity than constitutive promoters and known seed-specific promoters were discovered.

Growing community of inventors

Ibaraki, Japan

Fumio Takaiwa