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Candida glycerologenicum heat shock protein gene cgydj1 and its application

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

Active Publication Date: 2020-06-09
JIANGNAN UNIV
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  • Description
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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 cgydj1 and its application
  • Candida glycerologenicum heat shock protein gene cgydj1 and its application
  • Candida glycerologenicum heat shock protein gene cgydj1 and its application

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Embodiment

[0026] 1. C.glycerinogenes CCTCC M93018 gene CgYDJ1

[0027] The inventors selected S. cerevisiae by transforming the C. glycerinogenes CCTCC M93018 expressed genomic library into S. cerevisiae to screen for recombinant strains with improved heat resistance. After yeast plasmid extraction (yeast plasmid extraction kit purchased from BBI), nucleotide sequence sequencing (by Shanghai Bio Engineering Co., Ltd. completed sequencing) and obtained a large number of potential heat-resistant nucleotide sequences. Through nucleotide sequence and amino acid sequence alignment (by Basic Local Alignment Search Tool, clustalX2 and other analysis software), and structural analysis (by TMHMM-2.0, Swiss-model and other software), obtain the complete open reading frame of the gene , Amplify the fragment, the reaction system is as follows (50μl): 25μl Prime STAR Max Premix (2×), 15pmol primers, 150ng template, add double distilled water to 50μl (primers were synthesized by Shanghai Shenggong Bioen...

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Abstract

The invention belongs to the field of yeast gene engineering, and discloses a heat shock protein gene CgYDJ1 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 sequence table SEQ ID NO.1; more optimally, the DNA sequence is shown in the sequence table SEQ ID NO.1. The heat shock protein gene CgYDJ1 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 the C. glycerinogenes CCTCC M93018 gene CgYDJ1 of C. glycerinogenes CCTCC M93018 gene CgYDJ1 and its application. The gene CgYDJ1 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 infection. In order to improve the heat tolerance 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 directional modification of strains to improve their heat resistance. In yeast, the regu...

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