Development of growth selection systems to isolate a-type or α-type of yeast cells spontaneously emerging from MATa/α diploids

Journal of Biological Engineering

Shinya Honda¹  Nobuo Fukuda¹ 

[1] Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Higashi, Tsukuba, Ibaraki 305-8566, Japan

关键词: Promoters;    Transcription factors;    Gene regulation;    Biotechnology;    Yeast;   

Others  :  1139288
DOI  :  10.1186/1754-1611-7-27

 received in 2013-07-08, accepted in 2013-11-12,  发布年份 2013

【 摘 要 】

Background

Manufacture of MATa and MATα yeast cells is required for crossbreeding, a procedure that permits hybridization and the generation of new heterozygous strains. Crossbreeding also can be performed with a- and α-type of cells, which have the same mating abilities as MATa and MATα haploid cells, respectively.

Results

In this work, we describe a method to generate a- and α-type of cells via the naturally-occurring chromosomal aberration in parental MATa/α diploids. We successfully designed suitable genetic circuits for expression of the URA3 selection marker gene to permit isolation of a- and α-type of cells, respectively, on solid medium lacking uracil. Furthermore we succeeded in generation of zygotes by mating of both the manufactured a- and α-type of yeast cells.

Conclusions

This process does not require exposure to mutagens such as UV irradiation, thereby avoiding the accumulation of undesirable mutations that would detract from the valuable traits that are under study. All the genetic modifications in the current study were introduced into yeast cells using plasmids, meaning that these traits can be removed without altering the genome sequence. This approach provides a reliable and versatile tool for scientific research and industrial yeast crossbreeding.

【 授权许可】

   
2013 Fukuda and Honda; licensee BioMed Central Ltd.

【 图 表 】

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