Modification sequence of recombinant human mammilla tumor virus L1 capsid protein

A technology of human papillomavirus and capsid protein, which is applied in the direction of antiviral agents, viral antigen components, recombinant DNA technology, etc., can solve the problems of increased industrial cost, cumbersome production process, and increased consumption of specific reagents, so as to improve efficiency, The effect of reducing consumption and significant economic benefits

Inactive Publication Date: 2009-07-15
马润林 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the process of producing HPV L1 recombinant protein by genetic engineering methods, it is usually necessary to carry out anti-oxidation treatment on disulfide bonds, which increases the consumption of specific reagents and makes the production process cumbersome, resulting in increased industrial costs

Method used

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  • Modification sequence of recombinant human mammilla tumor virus L1 capsid protein
  • Modification sequence of recombinant human mammilla tumor virus L1 capsid protein
  • Modification sequence of recombinant human mammilla tumor virus L1 capsid protein

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Example 1 Taking HPV16 as an example to illustrate the modification of the amino acid sequence of HPV L1 capsid protein

[0032] 1. Detection and isolation of human papillomavirus type 16 DNA: The waste of clinical cell samples that may contain wild-type HPV16 virus (the remaining part of cervical exfoliated cells used for routine cytomorphological examination) was purchased from the Department of Gynecology, Beijing Anzhen Hospital clinic. Exfoliated cells (approximately 1 x 10 5 1) were treated with 1000 units of proteinase K (purchased from Huamei Biological Products Co., Ltd.) at 55 degrees Celsius for 300 minutes, and centrifuged at a high speed of 10000g for 30 minutes. The resulting supernatant could be used to detect whether there was HPV virus. The infected HPV16 positive cell supernatant can be directly used for PCR amplification of the target gene fragment of the HPV16 virus DNA sequence.

[0033] 2. Molecular cloning of HPV16 L1 protein DNA: HPV16 L1 capsi...

Embodiment 2

[0038] Example 2 Modification of the amino acid sequence of the L1 capsid protein of other common HPV types

[0039] The modification operation steps and detection methods of the amino acid sequence of the L1 capsid protein of other common HPV types are the same as those in Example 1, and the differences are shown in Table 1.

[0040] Table 1

[0041]

HPV

type

Primers used to clone the full length of the L1 protein gene

thing

Primers used for site-directed mutagenesis

used to replace

Cysteine

Acidic

amino acid HPV1 5'-ATGTATAATGTTTTTCAGATG-3'

5'-ATATACTAAGCCTTACGCCTGC-3' 5'-AGCCCCAGGGTGCAAATGATAG-3'

5'-GACTCTATTCATTTGCACCCTGG-3'

serine HPV2 5'-ATGCCAAGTTATGTCTTGTGG-3'

5'-GAGCTAACGCCTTACCCGTTTC-3' 5'-GGGGTTTGCTGCCCAGAATGAA-3'

5'-CACAGCTGTTCATTCTGGGCTG-3'

serine HPV3 5'-ATGGCACTGTGGCGCTCT-3'

5'-TGGCTATTTTTTGGTGCG-3' 5'-CGGTGTTCCTTTGCCCCAATG-3'

5'-ATTGGGGCAAAGGAACACCATG-3'

ser...

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Abstract

The invention relates to the prevention and treatment field of human papillomavirus infection, in particular to an amino acid modification sequence of recombinant human papillomavirus (HPV) L1 capsid protein, a nucleotide sequence encoding the amino acid modification sequence, and a carrier and a transformant containing the nucleotide sequence; and the invention further relates to application of an HPV L1 protein polymer consisting of the amino acid modification sequence to preparing vaccines, pharmaceutical compositions and diagnostic antigen or diagnostic antibodies. In the invention, an HPV L1 protein wild-type sequence is modified to prevent forming of a disulfide bond in the HPV L1 protein, the modification has no significant effect on the structural stability of the recombinant HPV L1 protein, but significantly improves the efficiency of purification process, and directly reduces the reagent consumption, thus effectively lowering the industrial production cost and having great economical benefit.

Description

technical field [0001] The present invention relates to the field of prevention and treatment of human papillomavirus infection. Specifically, the present invention relates to the amino acid modification sequence of recombinant human papillomavirus L1 capsid protein, the nucleotide sequence encoding the amino acid sequence, and the vector and transformant comprising these nucleotide sequences; The application of the HPV L1 protein multimer composed of the modified sequence in the preparation of vaccines, pharmaceutical compositions and diagnosis of antigens or antibodies. Background technique [0002] Papillomaviruses are a large group of DNA viruses that infect humans and other animals. Among them, human papilloma virus (HPV) is associated with various human diseases, including benign warts and cancers. [0003] Papillomavirus contains two structural proteins (or capsid proteins) for protecting viral chromosomes, namely L1 and L2. 360 L1 protein molecules or 72 L1 pentame...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/025C12N15/37C12N15/63A61K39/12A61K39/395A61P31/20
Inventor 马润林陈小江
Owner 马润林
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