Expression separation and purification method for Chinese Phytolacca acinosa cDNA mutant and TAT gene recombinant in prokaryon and application therefor

A recombinant and mutant technology, applied in DNA/RNA fragments, recombinant DNA technology, chemical instruments and methods, etc., can solve the problems of easy generation of drug resistance, large toxic and side effects, and achieve the effect of good biological activity
CN1952146AInactive Publication Date: 2007-04-25THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
Publication Date
2007-04-25
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention concerns a Ribosome inactibating protein (RIPS) Chinese mainland protein separated from plants which plays a role in anti-AIDS through catalyzing rRNA depurination of prokaryotic and eukaryotic ribosome with highly conserved stem-loop structure at the surface. The invention involves a Tat protein, the basic amino acid enrichment domain of which is protein transduction domain (PTD) and able to carry many exogenous protein directly into cells as protein orientation of cytokines or toxin. The invention involves a fusion protein of Tat and Chinese antiviral protein mutant. Performing deletion mutation to CAP using Quick-mutagenesis site-directed mutagenesis technology: 22 amino acid deletion in N-terminal, 29 amino acid deletion in C-terminal and site-specific mutagenesis: 151KI152 / 151AA152, 191FN192 / 191AA192. The recombinant protein of the invention has biological activity of anti-HIV-1 in vitro. It is found that CAP deletion and site-specific mutagenesis in fusion gene can significantly reduce its ability to combine with rRNA (cytotoxicity), however, without influence to its ability of combination of HIV RNA (anti-HIV activity), has more clinical application.
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Description

Technical field:

[0001] The present invention relates to two genes / proteins related to AIDS virus, the gene of Pokeweed antiviral protein and its encoded protein (CAP) mutant and a trans-acting activator (TAT) encoded by human immunodeficiency virus HIV-1 gene ) gene and its coded protein relate to the field of development and application of new gene and new protein in biomedical high technology. Background technique:

[0002] Acquired immunodeficiency syndrome (AIDS) is caused by the human immunodeficiency virus (HIV). Twenty years since the virus / disease was discovered, it has spread rapidly throughout the century, seriously threatening human health and social development. Since the first HIV patient was found in Peking Union Medical College Hospital on June 23, 1985, the prevalence of AIDS in my country has shown an accelerated upward trend in recent years. It is estimated that the actual number of HIV-infected people in my country has exceeded 600,000.

[0003] As one...

Claims

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