National Institute of Advanced Industrial Science and Technology

Micro Biopharm Japan Co., Ltd.

Tomohiro Tamura

Yoshikazu Fujii

Nobutaka Nakashima

Tadashi Fujii

Yasuo Mitani

Akira Arisawa

An expression vector capable of expressing a foreign gene in; a transformant ofproduced by using the expression vector; a method for producing a protein by using the transformant; a method for producing an active form of vitamin D3 from vitamin D3, which comprises highly expressing a gene encoding an enzyme involved in the synthesis of the active form of vitamin D3 in a transformant by using the expression vector or the transformant; a method for producing 25-hydroxyvitamin D2 from vitamin D2; and a method for producing pravastatin from compactin, which comprises highly expressing a compactin hydroxylase gene in a transformant by using the expression vector or the transformant.

Expression vector for pseudonocardia autotrophica

A vitamin Dhydroxylase is purified fromcell, and a primer is designed based on amino acid sequence obtained from hydroxylase. Subsequently, PCR is conducted using genomic DNA ofas a template to clone a gene for the vitamin Dhydroxylase. By conducting a conversion reaction using a microorganism in which the vitamin Dhydroxylase gene is expressed using a proper expression system, a hydroxide of vitamin D or the like (e.g., hydroxy vitamin D) can be produced with high efficiency.

Hydroxylase gene and use thereof

Provided is an expression vector capable of constitutive expression of a foreign gene in a bacterium belonging to the genus. The vector is a constitutive expression vector for a bacterium belonging to the genuscomprising: DNA comprising a nucleotide sequence of a mutated TipA gene promoter where a mutation is incorporated into a &#x2212;10 region sequence of a TipA gene promoter, the mutated TipA gene promoter being capable of thiostrepton-independent and constitutive expression of a gene located downstream thereof; a promoter sequence for the constitutive expression of a foreign gene; a ribosome-binding site sequence located downstream of the promoter sequence; and a multiple-cloning site sequence capable of incorporating a foreign gene therein, located downstream of the ribosome-binding site sequence.

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A microorganism of the genusis provided which has a higher sensitivity to lysozyme at a low concentration than a wild-type strain, which can easily cause cell lysis, and from which a recombinant protein expressed therein is easily recovered. More specifically, a mutant microorganism of the genushaving a higher sensitivity to lysozyme than a wild-type microorganism of the genus

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Sapporo, Japan

Tomohiro Tamura