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Method and identification method for improving the sulfur tolerance ability of grape juice yeast by transfecting ssu1 gene

A technology of grape juice yeast and grape juice, which is applied in the determination/inspection of microorganisms, biochemical equipment and methods, and the introduction of foreign genetic material using vectors, etc.

Inactive Publication Date: 2018-03-30
SOUTHWEST FORESTRY UNIVERSITY
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

However, in grape juice yeast, genes associated with sulfur tolerance SSU1 rarely reported

Method used

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  • Method and identification method for improving the sulfur tolerance ability of grape juice yeast by transfecting ssu1 gene
  • Method and identification method for improving the sulfur tolerance ability of grape juice yeast by transfecting ssu1 gene
  • Method and identification method for improving the sulfur tolerance ability of grape juice yeast by transfecting ssu1 gene

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Embodiment Construction

[0038] Materials: The non-sulfur-tolerant grape juice yeast strain is ACY338 and A4, the sulfur-tolerant grape juice yeast strain is A9, and the control Saccharomyces cerevisiae strain is EC1118. The above strains are all quoted from the University of Auckland, New Zealand (introduced by Zhang Hanyao, 2010), Escherichia coli DH5α, the above strains are stored in our laboratory. Grape juice yeast strains can be obtained from Professor Richard Gardner of the University of Auckland. Contact as below:

[0039] Wine Science Programme, School of Biological Sciences, University ofAuckland, Private Bag 92019, Auckland, New Zealand

[0040] Reagents: The primers were synthesized by Shanghai Bioengineering Company; Taq DNA polymerase, dNTPs, and RNase were purchased from Shanghai Bioengineering Company; DNA Marker is a 1Kb plus ladder, purchased from Invitrogen; Yeast genomic DNA extraction kit was purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd. Other conventional reagen...

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Abstract

The invention discloses a method for improving the sulfur tolerance ability of grape juice yeast by transfecting the SSU1 gene and an identification method, belonging to a method for cultivating new strains of yeast. The invention includes: extraction and purification of grape juice yeast DNA: construction of SSU1 gene expression vector; grape juice yeast transgene; determination of sulfur tolerance of grape juice yeast strains; PCR screening of transformants; sequencing of PCR products: fermentation of sulfur-containing grape juice after sterilization test etc. Sequencing showed that the candidate transformants all contained the characteristic sequence of the transgene. The test showed that both the transformants ACY330-SSU1 and A4-SSU1 could ferment well in the substrate containing 15 mg / L sodium sulfite. The present invention obtains the grape juice yeast sulfur-tolerant transgenic progeny strain, and the identification accuracy of the transformant strain is extremely high.

Description

Technical field [0001] The invention belongs to a yeast breeding and identification method. Background technique [0002] In the wine production process, a variety of antioxidants are needed to inhibit the growth of bacteria and ensure the quality and flavor of the wine. As the most common antioxidant, sulfur dioxide is widely used in winemaking processes. However, excessively high concentrations of sulfate dioxide ions in winemaking combine with free metal ions produced by grapes to form sulfites. Sulfites not only directly damage the tissue structure of cells, but also combine with some enzymes or metabolites to hinder cells. Normal metabolic activity. Therefore, the sulfite tolerance has also become one of the important indicators for screening brewing strains. [0003] Previous studies have shown that in the process of sulfur metabolism in Saccharomyces cerevisiae, genes related to sulfur tolerance SSU1 It can be used as a "sulfur pump" in cell regulation, thereby "improving...

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/81C12Q1/6895C12Q1/04
Inventor 张汉尧刘小珍张智铭贺笑
Owner SOUTHWEST FORESTRY UNIVERSITY