Method for constructing ribosome inactivating protein gene virus vector and expressing active proteins in tumor cell through ribosome inactivated protein gene virus vector

Abstract

The invention belongs to the technical field of bioengineering and specifically discloses a method for constructing a ribosome inactivating protein gene virus vector and expressing active proteins intumor cells through the ribosome inactivated protein gene virus vector. The method comprises the following steps that first, suitable ribosome inactivated proteins are selected, and codon optimizationfor humanized expression is conducted on mature region genes of the ribosome inactivated proteins; second, an adenovirus carrier of wild type and optimized genes is constructed, and recombinant viruses are packaged; and third, recombinant adenoviruses infect the tumor cells, the recombinant adenoviruses express proteins in the tumor cells through detection, and the anti-tumor effect of the recombinant adenoviruses is detected. According to the method, an alpha-charantin gene is subjected to codon optimization, an expression vector system for the adenoviruses capable of infecting the tumor cells of mammals is constructed so as to be suitable for expressing the active proteins in the tumor cells, and the defects that in the prior art, a prokaryotic cell expression system can only be expressed in escherichia coli when being used, and a plant virus vector can only be expressed in plant cells but cannot be expressed in the tumor cells and kill tumors when being used are overcome.

Description

Technical field: [0001] The application of the present invention belongs to the technical field of bioengineering, and more specifically relates to a method for constructing a ribosome inactivating protein gene viral vector and expressing the active protein in tumor cells. Background technique: [0002] Ribosome Inactivating Proteins (RIPs) are a class of toxic proteins mainly distributed in plants, which have RNA N-glycosidase activity. RIPs have highly conserved active cleft residues and secondary structures in the active site region, which can act on the large subunit 28S of eukaryotic cells to inactivate ribosomes, inhibit protein biosynthesis, and produce cytotoxic effects on cells. RIPs are divided into three types. Type I RIP is a single peptide chain protein with a molecular weight of about 11-30 kDa, such as bitter melon ribosome inactivating protein, trichosanthin (TCS), pokeweed protein (PAP), loofah toxin ( 1uffin), saporin, etc. Most ribosome inactivating prote...

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Problems solved by technology

[0004] However, whether the ribosome inactivating protein is expressed in Escherichia coli using a prokaryotic vector or expressed in a plant virus vector system, it cannot be expressed in mammalian cells and cannot be used for anti-tumor gene therapy

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