Targeted integration of neomycin into yeast artificial chromosomes (YACs) for transfection into mammalian cells | Zendy

J. Riley | Zendy; John Morten | Zendy; Rakesh Anand | Zendy

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Targeted integration of neomycin into yeast artificial chromosomes (YACs) for transfection into mammalian cells

Author(s) -

J. Riley,

John Morten,

Rakesh Anand

Publication year - 1992

Publication title -

nucleic acids research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 9.008

H-Index - 537

eISSN - 1362-4954

pISSN - 0305-1048

DOI - 10.1093/nar/20.12.2971

Subject(s) - biology , selectable marker , yeast artificial chromosome , plasmid , genetics , insert (composites) , human artificial chromosome , microbiology and biotechnology , homologous recombination , endonuclease , transfection , restriction enzyme , electroporation , gene , gene mapping , chromosome , mechanical engineering , engineering

Vectors have been constructed for the introduction of the neomycin resistance gene (neo) into the left arm, right arm or human insert DNA of yeast artificial chromosomes (YACs) by homologous recombination. These vectors contain a yeast selectable marker Lys-2, i.e. the alpha-aminoadipidate reductase gene, and a mammalian selection marker, neo, which confers G418 resistance. The vectors can be used to modify YACs in the most commonly used yeast strain for YAC library construction, AB1380. Specific targeting can be carried out by transfection of restriction endonuclease treated linear plasmids, with highly specific recombinogenic ends, into the YAC containing yeast cells. Analysis of targeted YACs confirmed that all three vectors can target correctly in yeast. Introduction of one of the targeted YACs into V79 (Chinese hamster fibroblast) cells showed complete and intact transfer of the YAC.

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