Agency of Industrial Science and Technology

Ministry of International Trade & Industry

Agency of Industrial Science and Technology Ministry of International

Showa Sangyo Co., Ltd.

Hidekatsu Maeda

Hideo Suzuki

Kazunari Taira

Aizo Yamauchi

Shinsuke Miyoshi

Hideaki Shinshi

Akira Kamibayashi

Hiromi Ishikawa

Hideoki Tanaka

Fumio Fukui

Kensuke Furukawa

Masanao Oda

Satoshi Nishikawa

Shigeru Kajiwara

Susumu Maruyama

The invention provides a recombinant plasmid containing a sequence encoding any genes inserted between 5' and 3' self-cleavage ribozymes. The recombinant plasmid can be amplified in vivo as well as in vitro while growing the host cell. When obtaining RNA transcripts of the inserted sequence, the recombinant plasmid does not require a restriction enzyme digestion step (run-off transcription) since cis-acting ribozymes perform self-catalyzed cleavage at 5' and 3' sides of the inserted sequence once it is transcribed. In this specific example, the trans-acting RNA enzyme sequence is inserted between 5' and 3' cleavage ribozymes. However, the trans-acting ribozyme sequence in the recombinant plasmid can be replaceable with any other sequence (e.g., antisense RNA, RNAs of HIV-1, HDV and other RNA viruses etc.). This construct is especially useful since each unit, consisting of 5' processing ribozyme, inserted sequence, and 3' processing ribozyme, can be connected in tandem. By so doing, ribozymes targeted to various sites can initially be transcribed as a long RNA chain which subsequently undergoes cleavage to produce independent trans-acting ribozymes, each possessing a specific target site.

RNA transcription system using novel ribozyme

A DNA fragment is disclosed which has connected to the downstream side of a first DNA sequence containing a region coding for the first ribozyme RNA a second DNA sequence containing a region coding for second ribozyme RNA capable of cleaving by self-processing the 3'-terminus site of the first ribozyme RNA subjected to transcription. A ribozyme having the 3'-terminus site thereof self-processed is obtained by effecting transcription of a RNA using as a template a recombinant vector obtained by recombination by the use of the DNA fragment.

Hammerhead ribozymes with enhanced stability provided by an additional 3' h

L-Tyr-L-Val, L-Leu-L-Tyr, L-Leu-L-Pro, L-Phe-L-Tyr, L-Tyr-L-Arg and its acid addition salts, L-Leu-L-Ser-L-Pro, L-Leu-L-Gln-L-Pro, L-Phe-L-Leu-L-Pro, L-Leu-L-Leu-L-Pro, L-Leu-L-Asn-L-Pro, L-Phe-L-Pro, L-Leu-L-Ala-L-Ala, L-Val-L-Ala-L-Ala, L-Leu-L-Gln-L-Gln, L-Val-L-Ala-L-Tyr, L-Leu-L-Ala-L-Tyr, L-Leu-L-Ser-L-His and its acid solution salts, L-Ile-L-Arg-L-Ala and its acid addition salts, L-Leu-L-Arg-L-Pro and its acid addition salts, and L-Ile-L-Arg-L-Ala-L-Gln-L-Gln and its acid addition salts, derived from a corn protein .alpha.-zein, and tripeptides represented by the formula

Oligopeptide, angiotensin converting enzyme inhibitors, hypotensive

An immobilized enzyme having an enzyme entrapped in gaps formed in a macromolecular gel matrix is produced by dispersing the enzyme in the form of a fine powder in an organic solvent having dissolved therein a polymerizable monomer, polymerizing the monomer thereby giving rise to a gel matrix, and displacing the organic solvent in the gel matrix with an aqueous solvent.

Enzyme immobilization by entrapment in a polymer gel matrix

Immobilized enzymes are produced by adding radioresistant enzymes to 5-25% aqueous solutions of polyvinyl type high molecular compounds and irradiating the enzyme-containing solutions with a high dosage of ionizing radiant rays in an inert atomsphere or under a decreased pressure for thereby gelling said solutions.

Method for production of immobilized enzyme

Growing community of inventors

Nagareyama, Japan

Hidekatsu Maeda