Toyo Boseki Kabushiki Kaisha

Cellfree Sciences Co., Ltd.

Biomerieus B.v.

Bunsei Kawakami

Yoshihisa Kawamura

Masao Kitabayashi

Tadayuki Imanaka

Masahiro Takagi

Toshihiro Kuroita

Masaaki Morikawa

Hiroaki Inoue

Taku Arakawa

Yoshiaki Nishiya

Hideyuki Komatsubara

Hideki Kamimura

Yoshihiko Maekawa

Yaeta Endo

Kenji Masuda

Shigeaki Nishii

Shigemichi Nishikawa

Akio Sugiyama

Yoshikazu Ishida

Yoichi Kadokami

Tomomi Yamazaki

Nobuhiro Katsuragi

The present invention provides an expression vector which is effective in an efficient establishment of transformed cells which express the aimed protein gene in a high level. An expression vector which has a cassette for expressing the drug selective marker gene containing mRNA destabilizing sequence, at least one element for stabilizing the gene expression and a cassette for expressing the gene of the aimed protein. Preferably, the mRNA destabilizing sequence is derived from AT-rich sequence existing in the 3'-untranslated region of cytokine, interleukin or proto-oncogene, and the element for stabilizing the gene expression is derived from Chinese hamster genome.

Expression vector for establishing highly productive cell and the highly productive cell

The present application relates to mutated RNA polymerases from bacteriophages that have increased stability, for example under high temperature conditions. Preferred mutated RNA polymerases according to the invention are mutant RNA polymerases from T7 or SP3 bacteriophages. An especially preferred embodiment of the present invention is a T7 RNA polymerase with a serine to proline amino acid change in the protein at position 633 of the amino acid sequence.

RNA polymerase mutants with increased thermostability

An object of the present invention is to provide a thermostable DNA polymerase with enhanced amplification efficiency and/or improved fidelity in polymerase chain reaction (PCR), and provide a process for production thereof. More specifically, the present invention provides thermostable DNA polymerase wherein in the DXEXXXH sequence (D: aspartic acid, E: glutamic acid, H: histidine, X, X, Xand X: any amino acid) consisting of DXE sequence within the EXO I region and a four amino acid length peptide adjacent to said glutamic acid(E) of thermostable DNA polymerase having 3'-5&#x2032; exonuclease activity, histidine(H) has been replaced by another amino acid.

Modified thermostable DNA polymerase

The present invention provides a composition for cell-free protein synthesis, which is superior in storage stability in a freeze-dried state, more particularly a freeze-dryable or freeze-dried composition for cell-free protein synthesis, which contains a cell extract for cell-free protein synthesis and inositol, and a freeze-dryable or freeze-dried composition for cell-free protein synthesis containing a cell extract for cell-free protein synthesis, and a deliquescent material in a proportion of not more than 0.01 part by weight per part by weight of a protein in the composition; and a composition for cell-free protein synthesis superior in storage stability in a frozen state, more particularly a freezable or frozen composition for cell-free protein synthesis, containing a cell extract for cell-free protein synthesis and polyhydric alcohol.

Composition for cell-free protein synthesis

A nucleic acid amplifying enzyme having a short reaction time and high fidelity is provided. The enzyme of this invention is a thermostable DNA polymerase having a nucleic acid extension rate of at least 30 bases per second and a 3&prime;-5&prime; exonuclease activity. Also provided are a method and kit for amplifying nucleic acid.

Kit for amplifying nucleic acid

An electrophoresis apparatus comprising an arched lid 2 covering an electrophoresis tank 1, a suction opening 4 set in one end of the lid 2 and an exhaust opening 7 set in the other end of the lid 2. On the outside of the suction opening 4 is set a fan 5. The fan 5 absorbs the outside air, which flows along the inner side of the lid 2 and is exhausted from the exhaust opening 7 together with the vapor generated during electrophoresis, thereby preventing clouding and bedewing of the inner surface of the lid. Consequently, the inside of the electrophoresis tank 1 can be safely observed without opening the transparent or semi-transparent lid 2 but through it.

Apparatus for electrophoresis

A nucleic acid amplifying enzyme having a short reaction time and high fidelity is provided. The enzyme of this invention is a thermostable DNA polymerase having a nucleic acid extension rate of at least 30 bases per second and a 3'-5' exonuclease activity. Also provided are a method and kit for amplifying nucleic acid.

Methods of amplification using a thermostable DNA polymerase from the

Thermostable DNA polymerase from a hyperthermophilic archaeon strain KOD1

A modified thermostable DNA polymerase having 5% or less of the 3'-5' exonuclease activity of the enzyme before modification and a DNA polymerase composition for amplifying nucleic acid, which comprises the modified thermostable DNA polymerase having 0 to 5% of the 3'-5' exonuclease activity of the enzyme before modification and an unmodified thermostable DNA polymerase having 3'-5' exonuclease activity or a modified thermostable DNA polymerase having 100 to 6% of the 3'-5' exonuclease activity of a thermostable DNA polymerase before modification; a method for amplifying nucleic acid by use of said modified thermostable polymerase or said DNA polymerase composition; and a reagent therefor.

Modified thermostable DNA polymerase derived from pyrococcus sp. KOD and

A method for isolating a ribonucleic acid, which comprises dissolution of a sample containing the ribonucleic acid, such as cells, in an acidic solution containing a lithium salt and a chaotropic agent, bringing the ribonucleic acid into contact with a nucleic acid-binding carrier such as silica particles, thereby to allow selective adsorption of the ribonucleic acid alone onto said carrier, and eluting the ribonucleic acid from the nucleic acid-bound carrier; a reagent therefor; and a method for producing a cDNA from the ribonucleic acid isolated by this method. According to the present invention, a high purity ribonucleic acid can be isolated quickly and safely from a sample containing the ribonucleic acid.

Method for isolating ribonucleic acid

A recombinant vector comprising an incorporated chromosome DNA fragment containing a PvuI restriction enconuclease gene derived from Proteus vulgaris, a host transformed with the recombinant vector and a method of producing PvuI restriction endonuclease characterized in that the transformed host is cultivated and PvuI restriction endonuclease is harvested from the resulting culture. Since the host transformed with the recombinant vector of the present invention produces PvuI alone, it is unnecessary to remove PvuII in the purification process for PvuI, and further, since its productivity for nonspecific DNase is lower in comparison with conventional producer bacteria, DNase can be removed easily, making the production of PvuI easy.

Growing community of inventors

Tsuruga, Japan

Bunsei Kawakami