Hybridized tRNA (transfer ribonucleic acid) and application thereof to glutathione peroxidase preparation

A technology of glutathione peroxide and hybridization, applied in the direction of DNA / RNA fragments, recombinant DNA technology, enzymes, etc., can solve the problems of cumbersome operation, lack of targeting, long cycle of simulated enzymes, etc., and achieve wide application Prospects, high-efficiency preparation methods, and the effect of facilitating gene manipulation
CN105018482AActive Publication Date: 2015-11-04JILIN UNIV
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
JILIN UNIV
Publication Date
2015-11-04

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Abstract

The invention discloses hybridized tRNA (transfer ribonucleic acid) and application thereof to glutathione peroxidase preparation and belongs to the field of biotechnology. The hybridized tRNA relates to sequences SEQ ID No: 1-16, consists of 90 basic groups, and not only can serve as substrate tRNA for selenocysteine synthesis, but also can be recognized by autologous elongation factor EF-Tu of escherichia coli. A hybridized tRNA gene is obtained by a gene synthesis method, then a GPX (glutathione peroxidase) gene is synthetized or amplified, the hybridized tRNA gene and the GPX gene are assembled on a secretory prokaryotic expression vector capable of expressing the hybridized tRNA, a TAG engineering strain is transformed and subjected to induced expression in presence of sodium selenite, and a catalytic group Sec of the GPX is introduced into a substrate binding site of protein in a way that conventional amino acids enter a peptide chain, so that the protein is given with high GPX activity. The hybridized tRNA and application thereof to glutathione peroxidase preparation have the advantages that the method is simple, and zymoprotein is high in activity, yield and stability, so that the problems that natural GPX is limited in source and instable in property are solved.
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Description

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a hybrid tRNA and its application in preparing high-activity selenium-containing glutathione peroxidase. Background technique

[0002] The substrate of selenium-containing glutathione peroxidase (GPX) is glutathione (GSH), and the catalytic group is selenocysteine ​​(Sec). In organisms, GPX together with superoxide dismutase (SOD) and catalase (CAT) constitute the body's antioxidant defense system. GPX plays an important role in this system. It uses the substrate GSH as a reducing agent to decompose hydrogen peroxide and various hydroperoxides in the body, so it can remove reactive oxygen species (ROS) in the body and prevent lipid peroxidation. Treat various diseases caused by active oxygen, such as aging, ultraviolet radiation, cardiovascular and cerebrovascular diseases, diabetes, tumors, etc. Different from other antioxidant enzymes, GPX can not only scavenge ROS, b...

Claims

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