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HPV18 E and E7 fusion gene mutant as well as related biological material and coding protein thereof

A technology of biological materials and transgenic animals, which can be applied in the fields of microorganisms, genetic engineering, plant genetic improvement, etc., and can solve the problems such as the risk of vaccines

Inactive Publication Date: 2014-04-23
BEIJING UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since HPV18E6 and E7 genes are oncogenes, it is potentially dangerous to directly apply them to vaccine design

Method used

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  • HPV18 E and E7 fusion gene mutant as well as related biological material and coding protein thereof
  • HPV18 E and E7 fusion gene mutant as well as related biological material and coding protein thereof
  • HPV18 E and E7 fusion gene mutant as well as related biological material and coding protein thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Embodiment 1, eliminate the carcinogenicity of wild-type HPV18E6E7 fusion protein and the preparation of wild-type HPV18E6E7 protein mutant and its coding gene highly expressed in mammalian cells

[0053] 1. Preparation of wild-type HPV18E6E7 protein mutant coding gene

[0054] In order to improve the expression of HPV18E6E7 gene in mammalian cells, especially human cells, the codon optimization of the wild-type HPV18E6E7 gene was carried out in this example to obtain the codon-optimized HPV18E6E7 fusion gene, named HPV18-E6E7-1 gene.

[0055] In order to eliminate the potential carcinogenicity of the wild-type HPV18E6E7 protein (named wHPV18-E6E7-1), the amino acid sequence encoded by the wHPV18-E6E7-1 gene includes the following site mutations: the 52nd and 138th amino acids of the E6 protein are respectively replaced by Leucine and cysteine ​​were mutated to glycine (L52G and C138G) and positions 27 and 98 of the E7 protein were mutated from cysteine ​​to glycine (C2...

Embodiment 2

[0090] Embodiment 2, the vaccine of prevention and treatment cervical cancer

[0091] This example provides two vaccines for the prevention and treatment of cervical cancer, named E6E7-3 and E6E7-4 respectively. The active ingredient of E6E7-3 is recombinant adenovirus rAd-HPV18omfE6E7-3. The active ingredient of E6E7-4 is recombinant adenovirus rAd-HPV18omfE6E7-4.

[0092] Recombinant adenovirus rAd-HPV18omfE6E7-3 is a recombinant adenovirus obtained by co-transfecting the recombinant vector containing the HPV18-E6E7-3 gene of Example 1 and the adenovirus backbone vector into adenovirus packaging cells; The recombinant vector of HPV18-E6E7-3 gene is the recombinant vector pDC315-HPV18-E6E7-3 obtained by inserting the HPV18-E6E7-3 gene of Example 1 into the multiple cloning site of pDC315. The adenovirus backbone vector is pBH Glox△E1\3cre.

[0093] The recombinant adenovirus rAd-HPV18omfE6E7-4 is a recombinant adenovirus obtained by co-transfecting the recombinant vector c...

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Abstract

The invention discloses an HPV18 E and E7 fusion gene mutant as well as a related biological material and coding protein thereof. The HPV18 E and E7 fusion gene mutant disclosed by the invention is DNA molecule represented as a) or b): a) coding sequence is DNA molecule (named as HPV18-E6E7-4 gene) represented on 15th-805th sites of SEQ ID No. 1; b) coding sequence is DNA molecule (named as HPV18-E6E7-3 gene) represented on 15th-806th sites of SEQ ID No. 2. The HPV18-E6E7-4 gene or HPV18-E6E7-3 gene not only has high expression on mammalian cell but also is free from oncogenicity; and the fusion gene mutant has biological safety, and is capable of effectively preventing and treating cervical cancer.

Description

technical field [0001] The present invention relates to HPV18E6 and E7 fusion gene mutants and related biological materials and encoded proteins. Background technique [0002] Human papillomaviruses (HPV) are a class of DNA viruses with strict host range and tissue specificity, which mainly infect human skin or mucosal epithelial cells, causing benign and malignant lesions at the infection site. In addition to causing various skin and mucous warts, HPV is more important because it is the main pathogenic factor of cervical cancer. According to the global cancer statistics report, it is estimated that there were 530,000 new cases of cervical cancer in the world in 2008, and 275,000 women died of the disease, and 85% of cervical cancers occurred in developing countries. Studies have shown that about 99.7% of cervical cancers are related to HPV infection. Traditional treatment methods for cervical cancer, such as surgery, radiotherapy, and chemotherapy, only have certain curat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/62C12N15/861C12N7/01C12N5/10C07K19/00A61K39/12A61P31/20A61P35/00
Inventor 周志祥赵晨曾毅李千千李泽琳
Owner BEIJING UNIV OF TECH
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