Recombinant L-251 protein with anti-inflammatory effect secreted by Japanese lamprey oral gland

A technology of L-251, oral glands, applied in the field of recombinant Japanese lamprey oral glands to secrete anti-inflammatory L-251 protein, which can solve problems such as tissue and cell damage

Inactive Publication Date: 2007-07-11
LIAONING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] On the one hand, the inflammatory response can remove foreign antigens or pathogenic microorganisms, but on the other hand, it can cause tissue and cell damage

Method used

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  • Recombinant L-251 protein with anti-inflammatory effect secreted by Japanese lamprey oral gland
  • Recombinant L-251 protein with anti-inflammatory effect secreted by Japanese lamprey oral gland
  • Recombinant L-251 protein with anti-inflammatory effect secreted by Japanese lamprey oral gland

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Experimental program
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Effect test

Embodiment Construction

[0051] 1. Extraction of total RNA: Trizol reagent from GIBCO BRL was used. details as follows:

[0052] (1) Take the Japanese lamprey oral gland and place it in liquid nitrogen to grind rapidly;

[0053] (2) Weigh 0.2g glandular tissue and secretion, add 1ml TRIZOL reagent to prepare homogenate, and incubate at 4°C for 5min;

[0054] (3) Add 0.2ml chloroform, close the lid tightly, shake vigorously for 15second, then place on ice for 5min;

[0055] (4) 4°C, 12000g, centrifuge for 15min;

[0056] (5) Transfer the upper aqueous phase to another centrifuge tube, add 0.5ml of isopropanol, and incubate on ice for 10 minutes;

[0057] (6) 4°C, 12000g, centrifuge for 15 minutes;

[0058] (7) Discard the supernatant, add 1ml of 75% ethanol to the precipitate (containing RNA) to wash, and vortex to mix;

[0059] (8) Centrifuge at 10,000 g for 5 min at 4°C to obtain RNA precipitates;

[0060] (9) After air drying, dissolve with appropriate amount of TE or RNase-free water for late...

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Abstract

The invention discloses a cDNA sequence and cloning of L-251 protein excreted by Japanese lamprey oral gland and amino acid sequence corresponding to cDNA and expression in the engineering bacteria as well as connecting protein of CD11a and CD11b beta2 integrated agent (CR3 receptor), which inhibits neutrophil leucocyte from transmitting from endothelium cell to inflammation point.

Description

technical field [0001] Identification of neutrophil inhibitory activity of recombinant Japanese lamprey oral gland secretion PR-1 protein L-251 belongs to the field of biotechnology, involving the sequence and cloning of the cDNA of the Japanese lamprey oral gland secretion L-251 protein cDNA, and the amino acids corresponding to the cDNA The sequence and its expression in Escherichia coli or Pichia pastoris, and the binding protein of L-251 protein as CD11a and CD11b β2 integrin can block the CD11b / CD18 and CD11a / CD18 integrin (CR3 receptor), thereby inhibiting the migration of neutrophils across endothelial cells to sites of inflammation. It is suggested that the lamprey, a parasitic organism, escapes the host's inflammatory response. In addition to viruses, worms and other species, this is a new species that has escaped the host's immune monitoring mechanism, and it is also a potential treatment and prevention caused by inflammatory responses. New drugs for immunopathologi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C07K14/435C12N15/63C12N1/21A61K38/17A61P31/00
Inventor 李庆伟吴毓白洁王继红
Owner LIAONING NORMAL UNIVERSITY
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