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Lacripin-18 gene fragment, encoded protein and preparation method of Lentinus edodes c91-3 strain

A kind of gene fragment, coding protein technology, applied in the field of Latcripin-18 gene fragment, coding protein and preparation field of Lentinus edodes C91-3 strain

Inactive Publication Date: 2016-01-06
DALIAN MEDICAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, so far there are no relevant reports on the extraction of gene fragments, encoded proteins and preparation methods that can induce tumor cell apoptosis and autophagy from Lentinus edodes C91-3 bacteria

Method used

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  • Lacripin-18 gene fragment, encoded protein and preparation method of Lentinus edodes c91-3 strain
  • Lacripin-18 gene fragment, encoded protein and preparation method of Lentinus edodes c91-3 strain
  • Lacripin-18 gene fragment, encoded protein and preparation method of Lentinus edodes c91-3 strain

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Embodiment Construction

[0034] a. Extract the mycelia total RNA of Lentinus edodes C91-3 strain:

[0035] Take the mycelium of Lentinus edodes C91-3 strain cultured for 18 days, add liquid nitrogen to the mortar and grind it thoroughly, add Trizol and let it stand at room temperature for 30 minutes, after the sample melts, continue to grind until the lysate is transparent, and let it stand at room temperature After 5 minutes, centrifuge at 12000r / min for 5min, transfer the supernatant to a new centrifuge tube, then add 1 / 5 volume of chloroform, shake gently, and centrifuge at 12000r / min at 4°C for 15min; absorb 0.5ml of the upper aqueous phase solution And transfer to another clean 1.5ml centrifuge tube, add an equal volume of isopropanol, invert and mix well, let stand at room temperature for 15min, centrifuge again at 4°C, 12000r / min for 10min, discard the supernatant, then add 1ml of 75% ethanol Washing and drying, and finally dissolving in DEPC-treated water, the total RNA of the mycelia of Lenti...

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Abstract

The present invention discloses a Latcripin-18 gene fragment of a mushroom C91-3 strain, encoding protein and a preparation method thereof, wherein the Latcripin-18 gene fragment has a nucleotide sequence represented by SEQIDNO:1, and the encoding protein has an amino acid sequence represented by SEQIDNO:2. The preparation method for the Latcripin-18 gene fragment comprises: extracting the total RNA of mycelium of the mushroom C91-3 strain; carrying out reverse transcription on the total RNA of the mycelium to synthesize cDNA; carrying out PCR amplification on the target gene, wherein the obtained cDNA is adopted as a template, and PCR reaction primers having two restriction enzyme cutting sites such as EcoR V and Xho I are adopted to carry out a PCR reaction; and recovering and purifying the DNA fragment.

Description

technical field [0001] The present invention relates to a kind of extract from mushroom C91-3 bacterial strain Latcripin-18 Gene fragment, encoded protein and preparation method, the encoded protein has PI3K-like structure domain, has the functions of inducing tumor cell apoptosis, autophagy, regulating cell cycle and anti-tumor. Background technique [0002] Fungi are rich in a variety of bioactive molecules, including ribosome-inactivating proteins, antifungal proteins, lectins, ubiquitin-like proteins, polysaccharides, and kinases. The anti-tumor components are unique in the treatment of tumors with the advantages of less adverse reactions and significant curative effects. Lentinus edodes fungus C91-3 belongs to Basidiomycetes Basidiomycetes Pleurotaceae. This strain has been preserved in the General Microbiology Center of China Committee for the Collection of Microbial Cultures in 2013, and the preservation number is CGMCCNo.7354. Lentinus edodes C91-3 has the effects ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/31C12N15/10C07K14/375
Inventor 黄敏张思若刘奔钟民涛伦永志张伟王晓丽李星云曹婧宁安红
Owner DALIAN MEDICAL UNIVERSITY
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