Latcripin-9 gene segment of shiitake mushroom C91-3 strain, coding protein, preparation method and application

A technology of gene fragment and encoded protein is applied in the fields of Latcripin-9 gene fragment, encoded protein, preparation and application of Lentinus edodes C91-3 strain, and can solve the problems of different effects of gene fragments and the like

Inactive Publication Date: 2016-02-24
DALIAN MEDICAL UNIVERSITY
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Problems solved by technology

Although there have been reports on the extraction of gene fragments, encoded proteins and preparation methods that can induce tumor cell apoptosis from Lentinus edodes C91-3 bacteria, the effects of different gene fragments are not the same, and more expression methods need to be developed urgently. The gene fragment of mushroom C91-3cDNA with strong protein efficacy, good targeting and industrial value

Method used

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  • Latcripin-9 gene segment of shiitake mushroom C91-3 strain, coding protein, preparation method and application
  • Latcripin-9 gene segment of shiitake mushroom C91-3 strain, coding protein, preparation method and application
  • Latcripin-9 gene segment of shiitake mushroom C91-3 strain, coding protein, preparation method and application

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Embodiment Construction

[0038] a. Extract the total RNA of mycelium of Lentinus edodes C91-3 strain:

[0039] Take the mycelium of Lentinus edodes C91-3 strain cultured in the culture medium for 18 days, add liquid nitrogen to the mortar and grind it thoroughly, add 1ml of Trizol and let it stand at room temperature for 30 minutes. After the sample is melted, continue to grind until the lysate is transparent, at room temperature After standing for 5 minutes, centrifuge at 12000r / min for 5min, transfer the supernatant to another centrifuge tube, then add 0.2ml of chloroform, and after gentle shaking, centrifuge at 4°C, 12000r / min for 15min; absorb the upper aqueous solution 0.5ml and transfer to another clean 1.5ml centrifuge tube, add an equal volume of isopropanol, mix upside down and let stand for 15min at room temperature, centrifuge again at 4°C, 12000r / min, 10min, discard the supernatant, and then add 75% Wash with 1ml of ethanol and dry, and finally dissolve it with DEPC treated water, and the tot...

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Abstract

The invention discloses a Latcripin-9 gene segment of a shiitake mushroom C91-3 strain, coding protein, a preparation method and application. A nucleotide sequence of the Latcripin-9 gene segment is shown as SEQ ID NO:1, and a nucleotide sequence of the coding protein is shown as SEQ ID NO:2. The preparation method of the Latcripin-9 gene segment includes following steps: extracting mycelium total RNA of the shiitake mushroom C91-3 strain; performing reverse transcription on the mycelium total RNA to synthesize cDNA; performing PCR amplification on a target gene; taking obtained cDNA as a template, and using a PCR primer with two restriction enzyme cutting sites EcoRI/XhoI for PCR; recycling and purifying a DNA segment.

Description

technical field [0001] The present invention relates to a kind of extract from mushroom C91-3 bacterial strain Latcripin-9 Gene fragment, encoded protein, preparation method and application. The encoded protein has the functions of inducing tumor cell apoptosis, regulating body immunity, regulating cell cycle, anti-tumor, scavenging free radicals and decolorizing dye wastewater. 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 CGM...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12N15/10C07K14/375C02F3/00A61K38/16A61P35/00A61P39/06
Inventor 黄敏阮文静钟民涛张伟刘奔李星云王晓丽曹婧宁安红伦永志
Owner DALIAN MEDICAL UNIVERSITY
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