Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1997-06-06
1999-02-02
Houtteman, Scott W.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
536 2432, 5303879, C12Q 168
Patent
active
058663380
ABSTRACT:
Human checkpoint huCDC34, huRAD9.sub.compA, and huRAD9.sub.compB cDNAs shown in FIGS. 1, 2, and 3. A method for isolating a human checkpoint cDNA that is capable of restoring growth at a restrictive temperature in a yeast test cell, wherein the yeast test cell comprises a genome having a first gene that forms a DNA strand break at a restrictive temperature and a second gene that fails to induce a cell cycle arrest in response to the DNA strand break, whereby the growth of the yeast test cell is inhibited at the restrictive temperature, the method comprising the steps of: obtaining a human cDNA library comprising a plurality of human cDNA clones; inserting the human cDNA clones individually into plasmid vectors comprising a selectable marker gene; transforming a culture of the yeast test cells with the plasmid vectors from the preceding step; selecting for yeast test cells transformed with the selectable marker gene; growing the selected transformants at the restrictive temperature and isolating a candidate transformant capable of growing at the restrictive temperature; and identifying the human cDNA carried by the candidate transformant as a human checkpoint cDNA by sequencing the human cDNA carried by the candidate transformant and determining that the human cDNA is less than 50% homologous with both the first gene and the second gene. Also yeast checkpoint RAD17, RAD24, MEC1, MEC2, and MEC3 cDNAs shown in FIGS. 4-8.
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Groudine Mark T.
Hartwell Leland H.
Plon Sharon E.
Weinert Ted A.
Arizona Board of Regents on Behalf of the University of Arizona
Fred Hutchinson Cancer Research Center
Houtteman Scott W.
University of Washington
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