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Lacripin-8 gene fragment, encoded protein, preparation method and application of mushroom c91-3 strain

A technology of gene fragments and strains, which is applied in the fields of Latcripin-8 gene fragment, encoded protein, preparation and application of Lentinus edodes C91-3 strain, can solve the problems of different effects of gene fragments and the like

Inactive Publication Date: 2019-07-12
DALIAN MEDICAL UNIVERSITY
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  • Claims
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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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  • Lacripin-8 gene fragment, encoded protein, preparation method and application of mushroom c91-3 strain
  • Lacripin-8 gene fragment, encoded protein, preparation method and application of mushroom c91-3 strain
  • Lacripin-8 gene fragment, encoded protein, preparation method and application of mushroom c91-3 strain

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Embodiment Construction

[0029] a. Extract total RNA from the mycelia of Lentinus edodes C91-3 strain:

[0030] Take the mycelium of Lentinus edodes C91-3 strain cultured 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 30min, after the sample melts, continue to grind until the lysate is transparent, at room temperature After standing still for 5min, centrifuge at 12000r / min for 5min, transfer the supernatant to another centrifuge tube, then add 0.2 ml chloroform, shake gently, centrifuge at 4˚C, 12000r / min for 15min; absorb the upper layer of water Transfer 0.5 ml of phase solution to another clean 1.5ml centrifuge tube, add an equal volume of isopropanol, invert and mix well, let stand at room temperature for 15 min, centrifuge again at 4˚C, 12000 r / min for 10 min, discard the supernatant, Then add 1 ml of 75% ethanol to wash and dry, and finally dissolve the water with DEPC treatment, the total RNA of the mycelium of ...

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Abstract

The invention discloses a Latcripin-8 gene fragment of a mushroom C91-3 strain, and a coded protein, a preparation method and application thereof. The nucleotide sequence of the Latcripin-8 gene fragment is as shown in SEQ ID No. 1, and the amino acid sequence of the coded protein is as shown in SEQ ID No. 2. The preparation method for the Latcripin-8 gene fragment comprises the following steps: extracting total RNAs of mycelia of the mushroom C91-3 strain; subjecting the total RNAs of the mycelia to inverse transcription so as to synthesize cDNAs; carrying out PCR amplification on a target gene: carrying out a PCR reaction on with the obtained cDNAs as a template and with a PCR reaction primer having two enzyme sites, i.e., BamHI and NotI; and recovering and purifying the obtained DNA fragment.

Description

technical field [0001] The invention relates to a Latcripin-8 gene fragment extracted from Lentinus edodes C91-3 strain, a method for preparing the encoded protein and its use. The encoded protein has the functions of inducing tumor cell apoptosis, regulating body immunity, regulating cell cycle, anti-tumor and It has the functions of 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 Basidiomycotina Basidiomycetes Pleurotaceae, which has been preserved in General Microbiology Center of China Committee for Microbial Culture Collection in 2013 with the preservation number CGMCC No.7...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/31C12N15/10C07K14/375A61K38/16A61P35/00A61P39/06
CPCA61K38/00C07K14/375C12Q1/686C12Q2525/131
Inventor 黄敏栾杭琪钟民涛张伟王晓丽李星云曹婧宁安红刘磊
Owner DALIAN MEDICAL UNIVERSITY
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