Method for rapidly detecting industrial saccharomyces pastorianus ester metabolism genes

A Pasteurella and gene technology, applied in the field of rapid detection of industrial Pasteurella ester metabolism genes, can solve the problem of low throughput, achieve strong specificity and sensitivity, improve typical flavor and consistency, and shorten experimental time Effect

Active Publication Date: 2014-03-12
TSINGTAO BREWERY
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Problems solved by technology

However, due to the limitation of low throughput of this method, a rapid and accurate technique is needed to simultaneously detect the expression of multiple ester metabolism-related genes in Pasteurella.

Method used

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  • Method for rapidly detecting industrial saccharomyces pastorianus ester metabolism genes
  • Method for rapidly detecting industrial saccharomyces pastorianus ester metabolism genes
  • Method for rapidly detecting industrial saccharomyces pastorianus ester metabolism genes

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Embodiment 1

[0051] A method for rapidly detecting ester metabolism genes of Pasteurella industrialis, including extraction of total RNA, preparation of cDNA templates by reverse transcription, multiplex PCR reaction, capillary electrophoresis, and product fragment analysis, aimed at genes related to ester metabolism of Pasteurella industrialis ATF1-Sc, ATF1-Sb, ATF2-Sc, ATF2--Sb, EEB1-Sc, EEB1-Sb, EHT1-Sc, EHT1-Sb, IAH1-Sc, IAH1-Sb, reverse transcription reaction to prepare cDNA template application primers are SEQ ID NO.2, SEQ ID NO.4, SEQ ID NO.6, SEQ ID NO.8, SEQ ID NO.10, SEQ ID NO.12, SEQ ID NO.14, SEQ ID NO.16, SEQ ID NO.18, SEQ ID NO.20, primers SEQ ID NO.1, SEQ ID NO.3, SEQ ID NO.5, SEQ ID NO.7, SEQ ID NO.9, SEQ ID NO. 11. SEQ ID NO.13, SEQ ID NO.15, SEQ ID NO.17, SEQ ID NO.19. It also includes industrial Pasteurella β-actin genes ACT1-Sc and ACT1-Sb as internal reference genes. The primers used to prepare cDNA templates in reverse transcription reactions are SEQ ID NO.22 and SEQ...

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Abstract

The invention relates to biotechnology, and particularly relates to a method for rapidly detecting industrial saccharomyces pastorianus ester metabolism genes. The method comprises the following steps: extracting total RNA (Ribose Nucleic Acid), preparing a cDNA (complementary DNA) template by virtue of reverse transcription reaction, performing multiplex-PCR ((Polymerase Chain Reaction), performing capillary electrophoresis, and analyzing product segments, wherein the cDNA template is prepared by virtue of reverse transcription reaction by aiming at industrial saccharomyces pastorianus ester type metabolism-related genes ATF1-Sc, ATF1-Sb, ATF2-Sc, ATF2-Sb, EEB1-Sc, EEB1-Sb, EHT1-Sc, EHT1-Sb, IAH1-Sc and IAH1-Sb. Compared with the conventional gene expressing quantitative analysis technology (fluorogenic quantitative PCR), the GeXP (multiplex-Gene Expression) quantitative analysis technology adopted in the method has the characteristics of stronger specificity, stronger sensitiveness, high automatic degree and the like, the reliability and repeatability of results are ensured, and the experiment time is shortened greatly. The method is in favor of controlling the metabolism and content of esters during fermentation under different production conditions and enhancing the typicality and consistency of beer flavor; besides, product characteristics are enabled to be remarkable and new products can be developed by means of rapidly adjusting the flavor of esters in beer.

Description

technical field [0001] The invention relates to biotechnology, in particular to a method for rapidly detecting ester metabolism genes of industrial Pasteurella yeast. Background technique [0002] Beer is a popular low-alcohol and nutritious beverage. Today, more than 90% of the global beer market is Lager type beer, so the brewer's yeast (commonly known as "Lager yeast" or "Pasteur's yeast") used to brew lager beer has been most widely used in the beer industry. Lager's yeast is different from Saccharomyces cerevisiae (S.cerevisiae), which was sequenced as a model organism in 1996. It is an allopolyploid strain with high chromosomal ploidy and complex chromosome structure. In addition to the Saccharomyces cerevisiae gene, it also carries the genetic information of S. bayanus, which has been named Saccharomyces pastorianus. Therefore, there are two homologous genes Sc-(S.cerevisiae) and Sb-(S.bayanus) for most of the genes in Pasteurella. However, compared with Saccharomy...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q2531/113C12Q2537/143C12Q2565/125
Inventor 樊伟董建军贺扬陈璐万秀娟陈嵘陈鹏赵玉祥张翠
Owner TSINGTAO BREWERY
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