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Method for detecting klenow fragment repetition position in saccharomyces cerevisiae chromosome

A technology for Saccharomyces cerevisiae and Saccharomyces cerevisiae strains is applied in the field of detecting the repetitive position of large segments of Saccharomyces cerevisiae chromosomes, which can solve the problems of cumbersome experimental data analysis work, and achieve the effects of short experimental period, rapid detection and simple operation.

Active Publication Date: 2016-06-01
TIANJIN UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

However, this method needs to accurately confirm the location of the site to be repaired, and the experimental data analysis work is very cumbersome; in actual research work, it is often not necessary to confirm the precise location of the wrong integration

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  • Method for detecting klenow fragment repetition position in saccharomyces cerevisiae chromosome
  • Method for detecting klenow fragment repetition position in saccharomyces cerevisiae chromosome

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Embodiment 1

[0049] 1. Culture medium preparation

[0050] The formula of SC-Lys-Met auxotrophic culture plate is synthetic yeast nitrogen source YNB6.7g / L, glucose 20g / L, default mixed amino acid powder of lysine and methionine 2g / L, agar powder 20g / L.

[0051] The formula of YPD liquid medium is yeast extract 10g / L, peptone 20g / L, glucose 20g / L.

[0052] The formula of SC+5-FOA medium is synthetic yeast nitrogen source YNB6.7g / L, glucose 20g / L, mixed amino acid powder 2g / L, 5-fluoroorotic acid 1g / L, agar powder 20g / L.

[0053] 2. Experimental method

[0054] 1. Sequence the entire genome of the tested Saccharomyces cerevisiae, and the results are as follows: figure 1 , showing that there is a small repeating region (multi-copy DNA1) and two large repeating regions (multi-copy DNA2) on chromosome V, and the chromosome is named "source chromosome V (synV)".

[0055] 2. Add GAL1 promoter and Kl.URA3 nutritional screening marker to the centromere of chromosome V of BY4742 strain by using ...

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Abstract

The invention relates to the field of molecular biology, in particular to a method for detecting a klenow fragment repetition position in a saccharomyces cerevisiae chromosome.By combining genomic sequencing with endoreduplication backcrossing, the position of repeated klenow fragment genome DNA in the chromosome is confirmed quickly.Through the endoreduplication backcrossing method, the position of the repeated klenow fragment genome DNA in the saccharomyces cerevisiae chromosome can be detected efficiently and quickly and repaired, repeated klenow fragment genome DNA is deleted, there is no need to precisely confirm the position of a klenow fragment repetition site or conduct cumbersome experimental data analysis on saccharomyces cerevisiae to be detected, operation is easy, and the experimental period is short.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a method for detecting the repeat position of a large segment of yeast chromosome. Background technique [0002] Saccharomyces cerevisiae, also known as baker's yeast or budding yeast. Saccharomyces cerevisiae is the yeast most widely related to humans, not only because it is traditionally used to make food such as bread and steamed bread and brew wine, but also used as a eukaryotic model organism in modern molecular and cell biology, and its role is equivalent to that of prokaryotic The model organism Escherichia coli. Yeast cells have high-efficiency homologous recombination characteristics (Ma, H., et al. Gene (1987), 58, 201-216), so they are widely used in the research of plasmid construction, gene knockout and even genome construction (Gibson, D.G. , et al. Proc. Natl. Acad. Sci. U.S.A (2008). 105, 20404-20409). The researchers of the artificially synthesized Saccharomyc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6869C12Q1/6895
Inventor 元英进谢泽雄李炳志
Owner TIANJIN UNIV
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