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Method for locating sequence position causing special growth phenotype of yeast

A yeast and phenotype technology, applied in the field of locating the sequence position that causes the special growth phenotype of yeast, can solve the problems of time-consuming and labor-intensive, and achieve the effect of avoiding time-consuming and labor-intensive

Active Publication Date: 2017-06-06
TIANJIN UNIV
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Problems solved by technology

This process requires the use of yeast micromanipulators to dissect a large number of tetrads and to verify the genotypes of thousands of individual spores one by one, which is a time-consuming and labor-intensive process.

Method used

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  • Method for locating sequence position causing special growth phenotype of yeast
  • Method for locating sequence position causing special growth phenotype of yeast

Examples

Experimental program
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Effect test

Embodiment 1

[0047] Example 1: Sequence mapping of semisynthetic yeast yYW0062

[0048] The semi-synthetic yeast strain yYW0062 (a semi-synthetic yeast strain obtained from wild-type S. BY4741 does not exist (see figure 2 ). The transformants produced during the integration of the synthetic module were collected to construct a transformant library, and the transformant library was copied onto the YPGE medium, and the growth of the transformants on the YPGE was observed after 2 days. Selective culture conditions were set according to the above-mentioned special growth phenotypes, and the transformants could be divided into two types under the condition of selection pressure: transformant pools that grew normally on YPGE, and transformant pools that had growth defects on YPGE. Use the PCR tags unique to the semi-synthetic yeast chromosome and the PCR tag specific to the wild-type yeast chromosome to design amplification primers for each functional gene of the two, and extract the mixed ge...

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Abstract

The invention relates to the technical field of functional genomics and discloses a method for locating the sequence position causing special growth phenotype of yeast. The method utilizes the sequence difference of functional genes of full synthesis yeast or partial synthesis yeast and wild type yeast, a synthesis amplification primer and a wild type amplification primer are used to conduct PCR (Polymerase Chain Reaction) on special growth phenotype yeast and normal growth phenotype yeast, the DNA of the normal growth phenotype yeast and the DNA of the special growth phenotype yeast are respectively locked according to the result, and the strip functional gene position cannot be amplified after the amplification primer is used to amplify, so that the overlapped functional gene position of the two yeasts are rapidly determined, the position is the sequence position causing the special growth phenotype of the yeast, and the time and labor consuming process of verifying the genotype of a single strain is avoided.

Description

technical field [0001] The invention relates to the technical field of functional genomics, in particular to a method for locating sequence positions that cause special growth phenotypes of yeast. Background technique [0002] With the popularization of DNA sequencing technology, the whole genome information of more and more species has been revealed. In addition, the development of related omics technologies such as transcriptome, proteome, and metabolome has enabled scientists to understand various complex metabolic pathways in biological cells. And a deeper understanding of the fine-tuning mechanism. In recent years, the continuous reduction in the cost of DNA synthesis and the continuous updating of large-fragment DNA assembly technology have enabled people to use synthetic biology methods to achieve rational design or reengineering of large-scale DNA, thereby achieving the optimization of the characteristics of microbial cells or using microorganisms as cells. The fact...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/6809C12Q2531/113
Inventor 元英进吴毅赵猛李炳志
Owner TIANJIN UNIV
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