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Latcripin-2 gene fragment of mushroom C91-3 strain, encoding protein and preparation method thereof

A technology for gene fragments and encoded proteins, applied in the field of Latcripin-2 gene fragments, encoded proteins and preparations of Lentinus edodes C91-3 strain

Inactive Publication Date: 2014-01-01
DALIAN MEDICAL UNIVERSITY
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  • Abstract
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  • Claims
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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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  • Latcripin-2 gene fragment of mushroom C91-3 strain, encoding protein and preparation method thereof
  • Latcripin-2 gene fragment of mushroom C91-3 strain, encoding protein and preparation method thereof
  • Latcripin-2 gene fragment of mushroom C91-3 strain, encoding protein and preparation method thereof

Examples

Experimental program
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Embodiment Construction

[0032] a. Extract total RNA from the mycelia of Lentinus edodes C91-3 strain:

[0033] 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, and centrifuge at 12000r / min at 4°C for 15min; absorb the upper aqueous solution 0.5 ml and transferred to another clean 1.5 ml 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, and Add 1 ml of 75% ethanol to wash and dry, and finally dissolve in DEPC-treated water, the total RNA of the mycelium of Lentinus ed...

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Abstract

The present invention discloses a Latcripin-2 gene fragment of a mushroom C91-3 strain, encoding protein and a preparation method thereof, wherein the Latcripin-2 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-2 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 BamH I 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-2 Gene fragment, encoded protein and preparation method, the encoded protein has Pkinase domain, and has the functions of inducing tumor cell apoptosis and autophagy, regulating immunity and anti-tumor, regulating cell cycle and the like. 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.7354. Lentinus edodes C91-3 has the...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N9/12A61P35/00C12N15/10
Inventor 黄敏杜忠娟钟民涛伦永志刘奔李星云王晓丽张伟曹婧宁安红
Owner DALIAN MEDICAL UNIVERSITY
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