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C20orf24 protein deletion mutant and application thereof

A technology for deletion of mutants and proteins, applied in the field of medicine, can solve the problems of limited related drugs, achieve broad use value, and inhibit the proliferation and growth of cancer cells

Active Publication Date: 2020-06-26
JINAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the relevant drugs targeting this Ras are still very limited

Method used

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  • C20orf24 protein deletion mutant and application thereof
  • C20orf24 protein deletion mutant and application thereof
  • C20orf24 protein deletion mutant and application thereof

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Embodiment 1

[0026] 1. Construction of pLVX-Puro-mCherry-HA-C20orf24 recombinant plasmid: construct pLVX-Puro-mCherry-HA-C20orf24 recombinant plasmid on the backbone of pLVX-IRES-ZsGreen1 vector (Clontech, Cat#632187), using homologous Recombination method, using C112-ClonExpress II One Step Cloning Kit kit (Novazyme, Cat#C112-01), insert the two gene fragments of C20orf24 and mCherry-HA into the pLVX-IRES-ZsGreen1 vector in the order of mCherry-HA-C20orf24 , as follows:

[0027] (1) Total RNA was extracted from human colorectal cancer HCT116 cells (purchased from the Cell Resource Center of Shanghai Academy of Biological Sciences, Chinese Academy of Sciences), and its reverse transcription product was used as a cDNA template; C20orf24 was obtained by PCR amplification experiment, gel recovery and purification (GenBank login No. NM_018840.5) gene fragment; PCR primers are C20orf24 UP and C20orf24 DW.

[0028] C20orf24 UP: 5'-ATGTACCCATACGATGTTCCAGATTACGCTAGCGGCGGGCGGCGG-3';

[0029] C20o...

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Abstract

The invention discloses a C20orf24 protein deletion mutant and application thereof. The C20orf24 protein deletion mutant is C20orf24 protein of which a Rab5ip structure domain is deleted, and the amino acid sequence of the mutant is MSGGRRKEEPPQPQLANGALKVSVWSKVLRSDAA WEDKDEFLDGVLYLYFSNYLQIDEEEYGGTWELTKEGFMTSFALFMVIWIIFYTAIHYD. The C20orf24 protein deletion mutant has a function of inhibiting cancer cell propagation growth, and has wide use value when being applied to preparation of cancer treatment medicines.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a C20orf24 protein deletion mutant and application thereof. Background technique [0002] Colorectal cancer is one of the most difficult diseases to treat in the world, characterized by high morbidity, high mortality and poor survival prognosis. The current mainstream treatment method is a combination of surgery and radiotherapy and chemotherapy. In the field of targeted therapy, the angiogenesis factor (VEGF) inhibitor bevacizumab and the epidermal growth factor receptor blocker panitumumab are widely used. , cetuximab and erlotinib. Nevertheless, the clinical curative effect is still unsatisfactory. The occurrence of chemotherapy resistance and toxic side effects affects the prognosis of cancer patients. It is imperative to further develop drugs targeting oncogenes. [0003] In colorectal cancer, the Ras / MAPK signaling pathway is one of the most active oncogenic ...

Claims

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Application Information

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IPC IPC(8): C07K14/47C12N15/12A61K38/17A61P35/00A61P35/02
CPCA61K38/00A61P35/00A61P35/02C07K14/47
Inventor 何庆瑜汪洋张静
Owner JINAN UNIVERSITY