Recombinant adenovirus for expression of novel tumour suppressor gene p53

A technology of recombinant adenovirus and tumor suppression, applied in gene therapy, antineoplastic drugs, genetic engineering, etc., can solve the problem of low activity

Inactive Publication Date: 2007-05-23
广州市凯诺生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, it has been proved that this activation activity is low, and further research is needed.

Method used

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  • Recombinant adenovirus for expression of novel tumour suppressor gene p53
  • Recombinant adenovirus for expression of novel tumour suppressor gene p53
  • Recombinant adenovirus for expression of novel tumour suppressor gene p53

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0017] 1. Formation of the R72 tumor suppressor P53: the method is to use gene mutation technology to mutate the codon ccc (P72) of the wild-type 72nd proline into the codon cgg (R72) of arginine. The mutated region forms a new site for the endonuclease smal1.

[0018] 1). Amplification of the fragment from the 5'-end Nco1 to the 72nd codon of the human wild-type tumor suppressor P53 gene:

[0019] Take the N'-terminal-Nco1-mutated region in the cDNA of the human wild-type P72 tumor suppressor P53 gene

[0020] The segment was used as a template for PCR amplification with artificially synthesized primers.

[0021] The PCR primers were designed as follows:

[0022] Primer 1:

[0023]N-terminal: 5'-gccttccgggtcactg aggagccg-3'

[0024] 30mer Nco1

[0025] Primer 2: ccg is the arginine codon

[0026] N-terminal: 5'-gaatgccagaggctgctccc gtggcccctgca-3'

[0027] 35mer

[0028] After tailing and adding adenine, the PCR amplification product was connected to the T-easy ve...

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Abstract

The invention discloses the expression structure of a recombinant adenovirus p53 for expressing anti-tumor gene, characterized in: (1) the 72nd amino acid of the wild p53 anti-tumor gene changes from praline to arginine, other amino acids components and their arranging sequences do not change, (2) splicing the p53 anti-cancer gene to the shuttle vector pDC315 strong promoter mMCV downstream at its N-terminal with incision enzyme Nhe1 and C-terminal with incision enzyme BamH1 fragment, and (3) the recombinant adenovirus is replication defective adenovirus displacing junction of adenovirus Ad5 with Ad35 junction. The new p53 anti-cancer gene adenovirus has strong infectivity, broad infection tissue / cell, strong expression of p53 anti-cancer gene, and product with strong apoptosis.

Description

technical field [0001] The invention relates to a method for preparing a recombinant adenovirus of a new human tumor suppressor gene p53. The prepared recombinant adenovirus expresses a novel tumor suppressor gene p53 protein with a stronger apoptosis function than wild-type P53, and is used for multiple treatment of cancer. Therefore, its technical field is related to life medicine and biotechnology. Background technique [0002] The P53 gene is the main member of the tumor suppressor gene family, and it is the gene most closely related to the occurrence of human tumors. The wild-type p53 gene plays an important role in maintaining normal cell growth and inhibiting the proliferation of malignant tumor cells. Various factors in the environment, such as ultraviolet rays, radiation and chemical substances, as well as certain metabolites produced by the body itself, can cause cellular DNA damage. Under normal physiological conditions, the DNA of these cells can be repaired o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/861C07K14/435A61K48/00A61K38/17A61P35/00
Inventor 王尚武朱雅南
Owner 广州市凯诺生物科技有限公司
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