Abstract: 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.

Abstract: 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 Bifidobacterium, 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 Bifidobacterium; a DNA encoding a secretory signal peptide; a DNA encoding a heterologous polypeptide; and a terminator DNA functioning in the bacterium of the genus Bifidobacterium, efficiently secretes the two types of heterologous peptides, i.e., two types of antibodies having anticancer effects, outside the bacterial cell.

Abstract: 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 Bifidobacterium, 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 genus Bifidobacterium capable of secreting a heterologous polypeptide. Means for attaining the object is a bacterium of the genus Bifidobacterium transformed by a vector having an expression cassette sequentially comprising a promoter DNA functioning in a bacterium of the genus Bifidobacterium, 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 Bifidobacterium; and capable of secreting the scFv antibody with high efficiency outside the bacterial cell.

Abstract: 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.

Abstract: [Object] To provide a gene transfer carrier that specifically accumulates only at a disease site even with systemic administration, has a high protein expression rate, and disappears from the disease site accompanying a cure, and a method for the treatment of an ischemic disease using same. [Solution means] The above-mentioned object has been accomplished by providing a gene transfer carrier formed from an anaerobic bacterium that can grow specifically at the site of an ischemic disease and can express at least one type of protein that is useful for the diagnosis or treatment of an ischemic disease, a pharmaceutical composition containing the gene transfer carrier, and a treatment method for an ischemic disease utilizing same.

Abstract: The present invention relates to a novel secretory signal, a novel plasmid containing the secretory signal, a transformed anaerobic bacterium transformed with said plasmid, a gene transfer carrier consisting of said anaerobic bacterium, and a pharmaceutical composition containing said carrier.

Abstract: To provide a gene transfer carrier that specifically accumulates only at a disease site even with systemic administration, has a high protein expression rate, and disappears from the disease site accompanying a cure, and a method for the treatment of an ischemic disease using same. The above-mentioned object has been accomplished by providing a gene transfer carrier formed from an anaerobic bacterium that can grow specifically at the site of an ischemic disease and can express at least one type of protein that is useful for the diagnosis or treatment of an ischemic disease, a pharmaceutical composition containing the gene transfer carrier, and a treatment method for an ischemic disease utilizing same.

Abstract: 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.

Abstract: The present invention provides a method for producing a cytosine deaminase (CD)-expressing, 5-fluorouracil (5-FU)-resistant microorganism which can grow in anaerobic tumor tissues, can express CD, and has a resistance to 5-FU at a concentration that is at least effective for antitumor activity. More specifically, the method is a method (A) comprising performing subculture or acclimation culture of a CD-expressing microorganism which can grow in anaerobic tumor tissues, in the presence of 5-fluorocytosine (5-FC), or a method (B) comprising (1) performing subculture or acclimation culture of a microorganism which can grow in anaerobic tumor tissues and does not express CD, in the presence of 5-FU to produce a 5-FU-resistant microorganism and (2) transforming the 5-FU-resistant microorganism by introducing a CD gene. The present invention also provides the CD-expressing, 5-FU-resistant microorganism and a pharmaceutical composition comprising the microorganism.

Abstract: The present invention relates to a secretory signal, a plasmid containing the secretory signal, a transformed anaerobic bacterium transformed with said plasmid, a gene transfer carrier consisting of said anaerobic bacterium, and a pharmaceutical composition containing said carrier.

Abstract: The present invention provides a therapeutic agent for anaerobic diseases such as solid tumor, comprising in combination a pharmaceutical composition for the treatment of an anaerobic disease containing the transformed anaerobic microorganism as an active component and a pharmaceutical composition containing as an active component an anaerobic microorganism colonization and growth enhancer for enhancing the specific colonization and proliferation of the anaerobic microorganism at an anaerobic disease site. Furthermore, the present invention provides to an anaerobic microorganism colonization and growth enhancer for enhancing colonization and growth of the transformed anaerobic microorganism at a disease site that is in an anaerobic environment.

Abstract: A conventional shuttle vector constructed by fusing an E. coli-derived plasmid and a transformant-derived plasmid functions in both E. coli and the transformant bacterium, and there exists no expression vector that functions only in a non-E. coli transformant. The present invention provides an plasmid expression vector comprising (1) a plasmid replication unit that functions in an anaerobic microorganism other than E. coli and (2) a protein expression unit formed from DNA coding for a protein having target activity and a DNA fragment containing a promoter and a terminator that function in the anaerobic microorganism. The expression vector of the present invention is capable of being replicated only in a transformant, eliminating the risk of the replication of the transformant gene in other pathogenic or aerobic bacterium, providing an extremely safe and reliable vector and gene transporter for therapeutic application.

Abstract: [Object] To provide a gene transfer carrier that specifically accumulates only at a disease site even with systemic administration, has a high protein expression rate, and disappears from the disease site accompanying a cure, and a method for the treatment of an ischemic disease using same. [Solution means] The above-mentioned object has been accomplished by providing a gene transfer carrier formed from an anaerobic bacterium that can grow specifically at the site of an ischemic disease and can express at least one type of protein that is useful for the diagnosis or treatment of an ischemic disease, a pharmaceutical composition containing the gene transfer carrier, and a treatment method for an ischemic disease utilizing same.

Abstract: A conventional shuttle vector constructed by fusing an E. coli-derived plasmid and a transformant-derived plasmid functions in both E. coli and the transformant bacterium, and there exists no expression vector that functions only in a non-E. coli transformant. The present invention provides an plasmid expression vector comprising (1) a plasmid replication unit that functions in an anaerobic microorganism other than E. coli and (2) a protein expression unit formed from DNA coding for a protein having target activity and a DNA fragment containing a promoter and a terminator that function in the anaerobic microorganism. The expression vector of the present invention is capable of being replicated only in a transformant, eliminating the risk of the replication of the transformant gene in other pathogenic or aerobic bacterium, providing an extremely safe and reliable vector and gene transporter for therapeutic application.

Abstract: The present invention provides an obligately anaerobic lactic acid bacterium having no risk of causing side effects in normal tissue and promising as a safe therapeutic agent and a gene transporter for a disease that is in an anaerobic environment such as a solid tumor, a preparation method therefor, and an expression vector useful in the preparation method. The obligately anaerobic lactic acid bacterium of the present invention has been artificially mutated from being facultatively anaerobic to being obligately anaerobic and, furthermore, is capable of being transformed by an expression vector having introduced thereinto a gene for expressing an active protein useful for the treatment of a disease that is in an anaerobic environment.

Abstract: The present invention provides a shuttle vector for a microorganism of the genus Bifidobacterium (BM) and Escherichia coli having a wide host range and a large copy number in BM and capable of highly expressing a desired protein when used as an expression vector; an expression vector capable of expressing a desired gene in BM by use of the shuttle vector; BM transformed with the expression vector; and an antitumor agent comprising the BM as an active ingredient. It comprises a pTB6-derived region portion comprising a replication origin (oriV)-repB region of pTB6 but not comprising MembB, MobA, OrfI, and oriT regions of pTB6 and an Escherichia coli-derived plasmid portion comprising a replication origin (Puc Ori) region of Escherichia coli but having deleted DNA encoding an N-terminal region of an ampicillin resistance gene (ampR) expression product ?-lactamase.

Abstract: The present invention relates to a novel secretory signal, a novel plasmid containing the secretory signal, a transformed anaerobic bacterium transformed with said plasmid, a gene transfer carrier consisting of said anaerobic bacterium, and a pharmaceutical composition containing said carrier.

Abstract: The present invention relates to a novel secretory signal, a novel plasmid containing the secretory signal, a transformed anaerobic bacterium transformed with said plasmid, a gene transfer carrier consisting of said anaerobic bacterium, and a pharmaceutical composition containing said carrier.

Abstract: The present invention relates to a novel secretory signal, a novel plasmid containing the secretory signal, a transformed anaerobic bacterium transformed with said plasmid, a gene transfer carrier consisting of said anaerobic bacterium, and a pharmaceutical composition containing said carrier.

Abstract: The present invention provides a therapeutic agent for anaerobic diseases such as solid tumor, comprising in combination a pharmaceutical composition for the treatment of an anaerobic disease containing the transformed anaerobic microorganism as an active component and a pharmaceutical composition containing as an active component an anaerobic microorganism colonization and growth enhancer for enhancing the specific colonization and proliferation of the anaerobic microorganism at an anaerobic disease site. Furthermore, the present invention provides to an anaerobic microorganism colonization and growth enhancer for enhancing colonization and growth of the transformed anaerobic microorganism at a disease site that is in an anaerobic environment.