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Candida glycerologenicum heat shock protein gene cghsp10 and its application

A Candida and gene technology, applied in the field of genetic engineering, can solve problems such as non-directionality and instability, and achieve the effects of improving heat tolerance, improving survival rate, and improving tolerance

Active Publication Date: 2020-02-18
JIANGNAN UNIV
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Problems solved by technology

In order to improve the heat resistance of yeast cells, traditional breeding techniques such as high-temperature domestication, natural selection, and mutation breeding have played an important role, but their shortcomings such as instability and non-directionality are obvious

Method used

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  • Candida glycerologenicum heat shock protein gene cghsp10 and its application
  • Candida glycerologenicum heat shock protein gene cghsp10 and its application
  • Candida glycerologenicum heat shock protein gene cghsp10 and its application

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Embodiment

[0024] 1. C. glycerinogenes CCTCC M93018 gene CgHsp10

[0025] The present inventor transformed S. cerevisiae according to the expression genome library of C. glycerinogenes CCTCC M93018 and screened recombinant strains with improved heat resistance, extracted yeast plasmid (the yeast plasmid extraction kit was purchased from BBI company), and sequenced the nucleotide sequence (provided by Shanghai Biological Technology Co., Ltd. Engineering Co., Ltd. completed the sequencing), and obtained a large number of potential thermotolerance-related nucleotide sequences. Through nucleotide sequence and amino acid sequence alignment (completed by Basic Local Alignment Search Tool, clustalX2 and other analysis software), and its structural analysis (completed by TMHMM-2.0, Swiss-model and other software), the complete open reading frame of the gene is obtained , amplify the fragment, the reaction system is as follows (50 μl): 25 μl Prime STAR Max Premix (2×), 15 pmol primers, 150 ng tem...

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Abstract

The invention belongs to the field of yeast gene engineering, and discloses a heat shock protein gene CgHsp10 of Candida glycerinogenes CCTCC M93018. The gene is over-expressed in Saccharomyces cerevisiae, and the heat tolerance of S.cerevisiae can be obviously improved. The similarity of the nucleotide sequence of the heat shock protein gene and the DNA sequence shown in the sequence table SEQ IDNO.1 is 75% or above, is more preferably 85% or above, and is further more preferably 95% or above, and the gene has the DNA sequence with the same function as the DNA sequence shown in the sequencetable SEQ ID NO.1; more optimally, the DNA sequence is shown in the sequence table SEQ ID NO.1. The heat shock protein gene CgHsp10 of Candida glycerinogenes CCTCC M93018 has an important effect on improving the heat tolerance of S.cerevisiae through a gene engineering technology.

Description

technical field [0001] The invention relates to CgHsp10 gene of C.glycerinogenes CCTCC M93018 and application thereof. The gene CgHsp10 is overexpressed in S.cerevisiae to improve the tolerance of S.cerevisiae to heat stress, and belongs to the technical field of genetic engineering. Background technique [0002] Improving the heat resistance of industrial yeast strains can effectively reduce the use of cooling water and energy consumption, reduce production costs, and effectively reduce the chance of bacterial infection. In order to improve the heat resistance of yeast cells, traditional breeding techniques such as high-temperature domestication, natural selection, and mutation breeding have played an important role, but their shortcomings such as instability and non-directionality are obvious. Modern genetic engineering technology plays an important role in the directed transformation of strains to improve their heat resistance. The regulation of heat shock response in ye...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/31C12N15/11C12N1/19C12R1/865
CPCC07K14/40
Inventor 诸葛斌孙祥祥杨飞陆信曜宗红宋健
Owner JIANGNAN UNIV
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