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Synthetic peptide for detecting human immunodeficiency viru-1 (HIV-1)

A technology of HIV-1, synthetic peptide, applied in the field of medical diagnosis

Pending Publication Date: 2018-12-14
DONGGUAN PENGZHI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

So far, there are no effective drugs and vaccines to treat and prevent AIDS, so HIV infection has become the most serious infectious disease and public health problem in the world today

Method used

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  • Synthetic peptide for detecting human immunodeficiency viru-1 (HIV-1)
  • Synthetic peptide for detecting human immunodeficiency viru-1 (HIV-1)
  • Synthetic peptide for detecting human immunodeficiency viru-1 (HIV-1)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Design of Recombinant Proteins

[0089] The gene sequence encoding SEQ ID NO: 1 was synthesized by gene synthesis, and X77 (X77 represents the 77th position of the sequence shown in SEQ ID NO: 1, the same below), X95, X101, X105, X106, X124, The amino acid at the X127 site was mutated,

[0090] The amino acid at position X77 is mutated from T to V or N or S, the amino acid at position X95 is mutated from Q to E, the amino acid at position X101 is mutated from N to E or D or Q, and the amino acid at position X105 is mutated from G to K or H or R, the amino acid at position X106 is mutated from L to E, the amino acid at position X124 is mutated from P to D or E, and the amino acid at position X127 is mutated from K to D, E or Q.

[0091] Construct mutant clones according to the above mutation directions, wherein the preferred scheme is that the preferred mutation schemes X77, X101, X105, X124, and X127 are respectively V, E, K, D, and D named HIV-Ag-1, respectively N, D,...

Embodiment 2

[0093] Design of Recombinant Proteins

[0094] On the basis of Example 1, the amino acid sequences X18, X95, X106, X123 and X130 in HIV-Ag-1 to HIV-Ag-9 were designed for mutation.

[0095] The amino acid at position X18 is mutated from R to S, the amino acid at position X95 is mutated from Q to E, the amino acid at position X106 is mutated from L to E, the amino acid at position X123 is mutated from L to K, and the amino acid at position X130 is mutated from N to K.

[0096] The new sequences were named HIV-Ag-10 to HIV-Ag-18, respectively. And constructed into pMD18-T vector. They were named pMD18-T-HIV-Ag-10 to pMD18-T-HIV-Ag-18 for subsequent amplification and preservation of nucleic acid fragments.

Embodiment 3

[0098] Construction, induced expression and purification of recombinant protein expression vector

[0099] Construction and induced expression of recombinant protein expression vectors: design upstream primers (with EcoR I restriction site) and downstream primers (with BamH I restriction site), to pMD18-T-HIV-Ag-0 to pMD18-T- HIV-Ag-18 is used as a template to amplify the target gene. After the target gene was purified, it was digested with EcoR I (TaKara, Cat. No.: 1010A) and BamH I restriction endonuclease (TaKara, Cat. No.: 1040A), and incubated at 37°C for 2 hours. The digested product was purified and ligated with the vector pET-28a that had undergone the same digestion, and incubated at 22°C for 2h and at 16°C for 2h. The ligation product was transformed into competent Escherichia coli BL21 (NEB (New England Biolabs), catalog number: C2530H) by heat shock method, spread on LB plates containing 50ug / ml Kan, and incubated at 37°C for 16h. Positive clones were picked and ...

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Abstract

The invention relates to the field of medical diagnosis, and particularly relates to a synthetic peptide for detecting human immunodeficiency viru-1 (HIV-1). The synthetic peptide is obtained by mutating an amino acid sequence shown in SEQ ID NO:1, so that an anti-HIV-1 antibody can be better detected, and potential "false negative" and "false positive" results can be better avoided.

Description

technical field [0001] The invention relates to the field of medical diagnosis, in particular to a synthetic peptide for detecting HIV-1. Background technique [0002] AIDS caused by Human Immunodeficiency Virus (HIV) has spread all over the world since it was discovered in the 1980s. According to the report of the World Health Organization, as of the end of 2016, there were approximately 36.7 million HIV-infected people worldwide. As of June 30, 2017, there were 718,000 HIV / AIDS patients in my country. HIV mainly attacks the helper T lymphocyte system of the human body, selectively invades cells with CD4 molecules, destroys the immune ability of the human body, causes the immune system to lose resistance, and is often combined with serious opportunistic infections, resulting in various diseases and cancers in the human body. It survives in the human body and develops to eventually lead to AIDS. So far, there are no effective drugs and vaccines to treat and prevent AIDS, ...

Claims

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Application Information

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IPC IPC(8): C07K14/16G01N33/68G01N33/569
CPCG01N33/56988G01N33/6854C07K14/005G01N2333/16G01N2469/20C12N2740/16022C07K14/001
Inventor 崔鹏何志强孟媛
Owner DONGGUAN PENGZHI BIOTECH CO LTD
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