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Latcripin-16 gene segment, encoding protein and preparation method of mushroom C91-3 strain

A technology for gene fragments and encoded proteins, applied in the field of Latcripin-16 gene fragments, encoded proteins and preparations of Lentinus edodes C91-3 strain

Inactive Publication Date: 2014-07-02
DALIAN MEDICAL UNIVERSITY
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  • Claims
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Problems solved by technology

However, so far, there are no relevant reports about extracting gene fragments, encoded proteins and preparation methods that can induce tumor cell apoptosis and regulate cell cycle from Lentinus edodes C91-3 bacteria

Method used

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  • Latcripin-16 gene segment, encoding protein and preparation method of mushroom C91-3 strain
  • Latcripin-16 gene segment, encoding protein and preparation method of mushroom C91-3 strain
  • Latcripin-16 gene segment, encoding protein and preparation method of mushroom C91-3 strain

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Experimental program
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Embodiment Construction

[0031] a. Extract total RNA from the mycelia of Lentinus edodes C91-3 strain:

[0032] 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 a new centrifuge tube, then add 0.2ml chloroform, shake gently, centrifuge at 4°C, 12000rpm, 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, mix it upside down and let stand at room temperature for 15min, centrifuge again at 4°C, 12000rpm for 10min, discard the supernatant, then add 1ml of 75% ethanol to wash and dry, and finally Treat water with DEPC to dissolve, total RNA of the mycelia of Lentinus edodes C91-3 stra...

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Abstract

The invention discloses a Latcripin-16 gene segment, an encoding protein and a preparation method of a mushroom C91-3 strain. The Latcripin-16 gene segment has a nucleotide sequence shown in SEQ ID NO:1; the encoding protein has an amino acid sequence shown in SED ID NO:2. The preparation method of the Latcripin-16 gene segment comprises the following steps: extracting mycelium total ribonucleic acid (RNA) of the mushroom C91-3 strain; carrying out reverse transcription on the mycelium total RNA to synthesize cDNA; carrying out PCR amplification on a target gene; taking obtained cDNA as a template, and carrying out polymerase chain reaction (PCR) by taking a PCR reaction primer with EcoRI and XhoI restriction enzyme cutting sites; and recovering and purifying the DNA segment.

Description

technical field [0001] The present invention relates to a kind of extract from mushroom C91-3 bacterial strain Latcripin-16 Gene fragment, encoded protein and preparation method, the encoded protein has endoribonuclease L-PSP domain, has the functions of inducing tumor cell apoptosis, regulating body immunity, 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 C91-3 is a fungus belonging to the Basidiomycotina Basidiomycetes Pleurotaceae. This strain has been preserved in the General Microbiology Center of the China Committee for the Collection of Microbial Cultures in 2013, and the preservation number is CGMCC No.7354. Len...

Claims

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Application Information

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IPC IPC(8): C12N15/31C07K14/375A61P35/00C12N15/10
Inventor 黄敏张伟钟民涛伦永志刘奔李星云王晓丽曹婧宁安红
Owner DALIAN MEDICAL UNIVERSITY
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