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Human nipple virus DNA inserted vaccine, its preparation method and application

A chimeric vaccine, nipple technology, applied in the biological field, can solve problems such as danger

Inactive Publication Date: 2006-08-30
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, due to the transformation activity of E7 protein itself, there is a certain risk when it is directly applied to the human body. Removing the transformation activity of E7 and retaining its antigenicity has become a prerequisite for vaccine development.

Method used

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  • Human nipple virus DNA inserted vaccine, its preparation method and application
  • Human nipple virus DNA inserted vaccine, its preparation method and application
  • Human nipple virus DNA inserted vaccine, its preparation method and application

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Embodiment Construction

[0093] Below in conjunction with accompanying drawing, the present invention is described in further detail:

[0094] 1. Acquisition and recovery of HPV16 E7 gene and heat shock protein gene fragments amplified by PCR

[0095]Add 5μl of 10×Buffer, 5μl of 10×dNTP (final concentration 200umol / L), 1μl of Primer 1 and 2 (about 30pmol), 1μl of template DNA (about 0.1ug), ddH 2 O 31μl and mix well, add TaqDNA polymerase 1U, paraffin oil 40μl. Cycle 30 times under the following conditions, 94°C for 1 min, 55°C for 1 min, 72°C for 1 min, and finally extend for 5 min. The amplified products were detected by 1.5% agarose gel electrophoresis. The 310bp DNA fragment of the amplified product was recovered through low melting point (60° C.) agarose.

[0096] The extracted plasmid pKs70 was double-digested with restriction endonucleases BamHI and HindIII, and HSP70 was recovered from low-melting point (60°C) agarose for ligation reaction.

[0097] 2. The connection reaction system is as ...

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Abstract

A DNA-chimeric human papilla virus vaccine for treating the cancer of uterine cervix is prepared through using the PCR site-directed mutation method to change the cysteine codor at the site No.91 in Zn figure region at the terminal C of human papilla virus vaccine to glycine codor, fusing the gene 70 of heat shock protein of tubercle bacillus with the terminal C of mutated gene E7, inserting the fused gene in the plasmid pc DNA 3.1(-) of eucaryotic expression carrier, enzyme severing and sequence analysis for verifying the correctness of configuration, and using the vaccine to immunize mouse for testing it.

Description

technical field [0001] The invention relates to biotechnology, more to a human papillomavirus (HPV) type 16 virus vaccine, and also to a preparation method of a human papillomavirus type 16 DNA therapeutic vaccine; meanwhile, it also relates to an application of the vaccine. Background technique [0002] Cervical cancer is one of the malignant tumors that seriously endanger women's health and life. There are about 500,000 new cases and 250,000 deaths in the world every year. It is the second leading cause of women's death, most of which are in developing countries, accounting for 80% % or so (Cain JM, Science, 2000, 288: 1753-1754). In 1977, German scholar Zur Hausen et al. discovered human papillomavirus (human papillomavirus, HPV) DNA from cervical cancer specimens, speculating that HPV infection is related to cervical cancer. Further molecular epidemiological evidence shows that HPV is closely related to various human cancers, especially cervic...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K39/12A61P35/00C12N15/34
Inventor 伍欣星熊金虎李辉王同祥赵旻魏芸丘小萍谭云
Owner WUHAN UNIV
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