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Antigen epitope minimum motif peptide of human papilloma virus 58 type E6 protein

A technology of human papillomavirus and motifs, which can be applied to peptides, instruments, analysis materials, etc., and can solve problems that have not been reported

Inactive Publication Date: 2010-04-21
GUANGDONG NANWAN PHARMA TECH CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the research on the epitope identification of HPV58 type E6 protein has not been reported so far

Method used

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  • Antigen epitope minimum motif peptide of human papilloma virus 58 type E6 protein
  • Antigen epitope minimum motif peptide of human papilloma virus 58 type E6 protein
  • Antigen epitope minimum motif peptide of human papilloma virus 58 type E6 protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1: human papillomavirus type 58 (HPV58) E6 protein linear BCE scanning mapping

[0026] Materials and methods:

[0027] 1. HPV58 gene clone (pLink322 / HPV58 plasmid) was purchased from Dr. Matsukura T, Japan. The pRSET C plasmid was purchased from Invitrogen, USA. The heat-inducible expression plasmid pXXGST-1 was constructed by the inventor of this patent (patent application number: 200710173305.2). Escherichia coli BL21 (DE3) strain was preserved by the State Key Laboratory of Genetic Engineering, Fudan University.

[0028] 2. Restriction enzymes EcoR I, BamH I, Sal I, Taq enzyme and T4 DNA ligase were purchased from Japan TaKaRa Biotechnology Company, pre-stained protein molecular weight standards, horseradish peroxidase-labeled goat anti-rabbit secondary antibody (IgG / HRP ), diaminobenzidine (DAB) and 0.2 μm nitrocellulose membrane were purchased from Shanghai Sangon Biotechnology Service Company. 6×His monoclonal antibody was purchased from Shanghai R...

Embodiment 2

[0040] Example 2: Human papillomavirus type 58 (HPV58) E6 protein P14 epitope peptide minimum motif identification

[0041] Materials and methods:

[0042] See the corresponding part of Example 1.

[0043] The specific steps for identifying the minimal motif of the P14 epitope peptide are as follows:

[0044] 1. According to the results of E6 protein antigen epitope scanning and positioning in Example 1, the minimum motif identification was carried out by taking the E6-2 epitope peptide P14 as an example. Design a series of 8-peptide-encoded positive and negative strand fragments of 15 peptide sequences overlapping each other by 7 amino acid residues across E6-2 (positive strand 5'-end plus 5'-gatcc, 3'-end plus taag-3'; negative Add 5'-tcgactta to the 5'-end of the chain, add g-3' to the 3'-end), and send out DNA synthesis.

[0045] The 2-4 operation steps are the same as the 2-4 steps in the embodiment 1.

[0046] 5. The blotted membrane was repeatedly washed four times wi...

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Abstract

The invention belongs to the technical field of biological medicine and biological detection and particularly relates to antigen epitope minimum motif peptide of a human papilloma virus (HPV) 58 type E6 protein and an application thereof. The invention provides four epitope peptides (comprising an 8-peptide sequence which can be expressed in a fusion way with carrier proteins such as truncated GST188 and the like and contain epitope minimum motif) which can be used as antigens for specifically detecting HPV 58 type virus infection in a single way or a combination way and contains the minimum motif and a marker minimum motif peptide amino acid sequence used for specifically detecting low risk type HPV virus infection and medium-high risk type HPV virus infection in a discrimination way. The amino acid sequences of the minimum motif peptides and short peptides are shown as SEQ ID No.1-SEQ ID No.9.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and biological detection, and in particular relates to all linear B cell epitopes (B cell epitope, BCE, Or called antigenic determinant) minimal motif peptide and its extended 8-peptide, and the application of these motif peptides and short peptides. Background technique [0002] Human papillomavirus (human papillomavirus, HPV) is a general term for a group of viruses. At present, more than 100 types of HPV have been found, and about 35 types of them can infect the female genital tract epithelium, and the middle and high-risk HPV types closely related to cervical cancer and cervical intraepithelial lesions involve HPV16, 18, 31, and 33 , 45, 52, 56 and 58 and more than 20 types (Munoz N. human papillomavirus and cancer: the epidemiological evidence. J Clin Virol, 19: 1-5, 2000). It is clear that HPV viruses are very small, and its genome of about 8kb in length usually encodes 6 early protein...

Claims

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Application Information

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IPC IPC(8): C07K5/103C07K7/06G01N33/569
Inventor 徐万祥季朝能何亚萍唐海平顾少华王健孙志达谢毅
Owner GUANGDONG NANWAN PHARMA TECH CO LTD
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