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Application of gene YEL013W and gene YER042W

A gene and genome technology, applied in the direction of microorganism-based methods, biochemical equipment and methods, enzymes, etc., can solve the problems of natural genome rearrangement and mutation time span, low controllability of functional directed evolution, etc., to achieve increased yield Effect

Active Publication Date: 2018-08-03
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The time span of natural genome rearrangement and mutation is large, and the controllability of functional directed evolution is low

Method used

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  • Application of gene YEL013W and gene YER042W

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1: Construction of Saccharomyces cerevisiae strains producing carotenoids

[0027] Artificially synthesize three exogenous genes, crtE, crtI and crtYB, construct the heterologous carotenoid synthesis pathway crtE+crtI+crtYB, and then use the yeast homologous recombination mechanism to design the left and right connecting elements, respectively, and heterologous carotenoids The enzyme synthesis pathway was digested together and the gel was recovered, and the haploid synthetic synV chromosome Saccharomyces cerevisiae was co-transformed. The carotenoid pathway was integrated into the YEL063C / CAN1 locus in synV together with the Leu2 nutritional marker to obtain yeast strain yJBH000.

Embodiment 2

[0028] Example 2: Gene knockout test verification effect

[0029] The three genes YEL013W, YEL014C (for control) and YER042W in yJBH000 were knocked out individually and in combination to obtain yJBN009 (knockout YEL013W gene), yJBN010 (knockout YEL014W gene, yJBN006 (knockout YEL014C and YEL013W gene) gene), yJBN007 (knockout YER042W gene) and yJBN008 (knockout YEL014C, YEL013W and YER042W gene) five test strains. Then ferment production according to the following method:

[0030] The strain was inoculated into 5mL YPD medium for activation and cultured for 24 hours, then inoculated with 40mL of YPD medium (40g / L glucose) at an OD600 of 0.1, and fermented in a shaker at 30°C and 250rpm for 60 hours.

[0031] 2 mL of the fermentation culture was collected by centrifugation, and the carotenoids were extracted from the pellet with 1 mL of acetone. Samples were analyzed by HPLC. The mobile phase consisted of methanol-acetonitrile-dichloromethane (9:40:1 v / v), and the flow rate ...

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Abstract

The invention relates to the technical field of biology and particularly discloses a novel application of gene YEL013W and gene YER042W in a yeast genome. Yield of a yeast strain for producing carotenoids can be increased remarkably by knock out the two genes, and a specific gene target is provided for modification of microbial engineering bacteria.

Description

technical field [0001] The invention relates to the field of biotechnology, and more specifically relates to the application of YEL013W gene and YER042W gene. Background technique [0002] In recent years, with the continuous improvement of genome sequencing capabilities, it has been possible to resolve ubiquitous genome rearrangements that were difficult to find in the past. The main forms of genomic variation discovered in the 20th century are single nucleotide polymorphism (SNP) and heterochromatin polymorphism (HP). However, in recent years, studies have found that genome rearrangements are commonly found in microorganisms (such as Yersinia, yeast), plants (such as grass, rice), animals (such as lampreys, mice), and human genomes , in particular, a large number of structural variations (SVs) in the genome (including fragment deletions, duplications, horizontal migration, reciprocal translocations, inversions, etc.) Driving the evolution of life provides key evidence. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12P23/00C12R1/865
CPCC07K14/39C12N9/0051C12P23/00C12Y108/04
Inventor 元英进贾斌潘硕吴毅李炳志
Owner TIANJIN UNIV
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