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A method for repairing structural abnormality of yeast chromosome

A chromosome and yeast technology, applied in the field of genome engineering, can solve problems such as low transformation efficiency, ineffective repair of DNA, and small scale

Active Publication Date: 2020-07-28
TIANJIN UNIV
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Problems solved by technology

Existing chromosomal repair technologies involve relatively small repair DNA scales. For example, the use of CRISR-Cas9 technology can better realize the repair of point mutations or combine with homologous recombination technology to achieve the repair of DNA sequences with a length of several Kb. The synthesis of exogenous long DNA fragments and low transformation efficiency make CRISPR-Cas9 technology generally limited to changes in DNA sequences of a few Kb in length, and cannot effectively repair DNA of hundreds of Kb in length
At present, there are few reports on DNA repair technology for hundreds of Kb in length

Method used

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  • A method for repairing structural abnormality of yeast chromosome
  • A method for repairing structural abnormality of yeast chromosome
  • A method for repairing structural abnormality of yeast chromosome

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Embodiment 1

[0035] Example 1: The fully synthetic chromosome Saccharomyces cerevisiae R-2 repairing abnormal chromosome structure

[0036] Karyotype analysis of Saccharomyces cerevisiae R-2 by pulsed field gel electrophoresis image 3 It can be seen that compared with the wild-type control strain BY4741 in strain R-2, the band corresponding to chromosome 10 has obviously migrated upward. After analysis of strain R-2 by whole genome sequencing, by Figure 4 The sequencing depth map revealed that there were two regions with large fragment duplications. One mainly occurred in the megachunk C region, involving a sequence length of 101kb, which was a continuous three-copy duplication. The second mainly occurs in the megachunk E region, involving a sequence length of 147kb, which is two-copy and four-copy staggered duplication. The comprehensive calculation shows that the length of chromosome 10 of strain R-2 is 171kb longer than that of the wild type. Primers were designed for the sequence...

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Abstract

The invention relates to the field of genome engineering and discloses a method for repairing the anomaly of a yeast chromosome structure. According to the method disclosed by the invention, yeast chromosomes, which have chain exchange at a design region, are rapidly screened by utilizing an efficient homologous recombination mechanism and a screening label of yeast, and exchange between an sequence with the abnormal yeast chromosome structure and a normal chromosome sequence is realized, so that the aim of efficiently repairing the variation of the yeast chromosome structure in a large scale is realized.

Description

technical field [0001] The invention relates to the field of genome engineering, in particular to a method for repairing structural abnormalities of yeast chromosomes. Background technique [0002] Chromosomal structural abnormalities refer to abnormal chromosomal breakage, recombination or exchange resulting in chromosomal aberrations. The types of abnormal chromosome structure can be divided into DNA copy number variation, large insertion, large duplication, large turnover, and chromosomal translocation. Chromosomal structural abnormalities occur widely in most species, and many chromosomal structural abnormalities are closely related to cell aging, chromosomal genetic diseases, cancer and other diseases. For example, cat-calling syndrome is a genetic disease caused by the partial deletion of chromosome 5 in humans, and the notched wing of fruit flies is also caused by the deletion of a section of chromosome; the rod eye phenomenon of fruit flies is caused by the duplicat...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/90C12N1/19C12R1/865
CPCC12N1/18C12N15/90
Inventor 元英进吴毅李炳志
Owner TIANJIN UNIV
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