Transformation of yeast using calcium alginate microbeads with surface-immobilized chromosomal DNA | Zendy

Atsushi Mizukami | Zendy; Eiji Nagamori | Zendy; Yukiko Takakura | Zendy; Sachihiro Matsunaga | Zendy; Yoshinobu Kaneko | Zendy; Shin’ichiro Kajiyama | Zendy; Satoshi Harashima | Zendy; Akio Kobayashi | Zendy; Kiichi Fukui | Zendy

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Transformation of yeast using calcium alginate microbeads with surface-immobilized chromosomal DNA

Author(s) -

Atsushi Mizukami,

Eiji Nagamori,

Yukiko Takakura,

Sachihiro Matsunaga,

Yoshinobu Kaneko,

Shin’ichiro Kajiyama,

Satoshi Harashima,

Akio Kobayashi,

Kiichi Fukui

Publication year - 2003

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/03354st03

Subject(s) - yeast artificial chromosome , dna , microbead (research) , transformation (genetics) , in vitro recombination , molecular cloning , yeast , chromosome , biology , microbiology and biotechnology , cloning (programming) , chemistry , genetics , gene , gene mapping , complementary dna , computer science , programming language

Yeast artificial chromosomes (YACs) are useful cloning vectors that have the capacity to carry large DNA inserts. The largest barrier to using such large DNA molecules in transformation experiments has been their physical instability in solution. We developed a new method of transforming yeast using chromosome-sized DNA. The method uses calcium alginate microbeads to immobilize high-density yeast chromosomal DNA. Chromosomal DNA immobilized on microbeads is physically stabilized compared with naked chromosomal DNA. The microbead-mediated transformation performed well, not only with respect to the transformation frequency with large DNA molecules (> 100 kb) but also in successful transformation using split chromosome DNA that exceeded 450 kb.

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