Anaeropharma Science, Inc.

Shinshu University

Azusapharma Sciences, Inc.

Yuko Shimatani

Takashi Yano

Takayuki Sasaki

Takeshi Masaki

Satoshi Kobayashi

Hitomi Shimizu

Koichi Koseki

Koichiro Shioya

Tomio Matsumura

Jun Amano

Shun'ichiro Taniguchi

Minoru Fujimori

Yoshinori Hamaji

Yuko Wada

Masao Tsukamoto

Hitomi Matsuhashi

Kengo Inoue

Masami Okabe

Provided are: a transformation plasmid for transforming anaerobes and enabling highly efficient and stable secretory expression of a target protein; a gene delivery carrier formed from said anaerobes transformed by said plasmid; a pharmaceutical composition including said gene delivery carrier; and a method for diagnosing or treating an ischemic disease utilizing these. Also provided are: a novel secretory signal; a transformation plasmid including said secretory signal; a gene delivery carrier formed from anaerobes transformed by said plasmid; a pharmaceutical composition including said gene delivery carrier; and a method for diagnosing or treating an ischemic disease utilizing these.

Therapeutic agent for ischemic diseases

The object of the invention is to provide an anaerobic enterobacterium having a higher therapeutic effect on an anaerobic site such as a solid tumor tissue and an ischemic disease site. A bacterium of the genus, which is transformed with a plasmid co-expressing two types of heterologous polypeptides and comprising two types of secretory expression cassettes each sequentially comprising a promoter DNA functioning in the bacterium of the genus; a DNA encoding a secretory signal peptide; a DNA encoding a heterologous polypeptide; and a terminator DNA functioning in the bacterium of the genus, efficiently secretes the two types of heterologous peptides, i.e., two types of antibodies having anticancer effects, outside the bacterial cell.

Co-expression plasmid

An object is to provide a signal peptide that can secrete a heterologous polypeptide with high efficiency outside the bacterial cell in a bacterium of the genus, an expression cassette that can secrete a heterologous polypeptide with high efficiency outside the bacterial cell, a heterologous polypeptide expression vector, a bacterium of the genuscapable of secreting a heterologous polypeptide. Means for attaining the object is a bacterium of the genustransformed by a vector having an expression cassette sequentially comprising a promoter DNA functioning in a bacterium of the genus, a DNA encoding the secretory signal peptide, a DNA encoding a scFv antibody having an antitumor activity, and a terminator DNA functioning in the bacterium of the genus; and capable of secreting the scFv antibody with high efficiency outside the bacterial cell.

Heterologous polypeptide expression cassette

An object of the present invention is to provide a method of efficiently constructing a gene delivery carrier having a favorable activity and expression efficiency of a protein expressed by a gene introduced by transformation. Moreover, an object of the present invention is to provide a pharmaceutical composition comprising a gene delivery carrier constructed by the construction method and a therapeutic agent for solid tumor comprising the resistant bacterium. The objects of the present invention have been attained by a gene delivery carrier consisting of an anaerobic microorganism capable of growing in a tumor tissue in an anaerobic environment and capable of expressing (A) a protein having an antitumor activity or (B) a protein having an activity of converting an antitumor substance precursor to an antitumor substance, wherein DNA encoding the protein of interest is allowed to further comprise, on the 5'-terminal side thereof, a DNA fragment comprising a nucleotide sequence encoding a peptide consisting of an amino acid sequence from the 1st to at least 4th amino acids at the N-terminus of a histone-like DNA-binding protein.

Method of constructing gene transport support

Growing community of inventors

Nagano, Japan

Yuko Shimatani