Abstract
Human specific "integrative selection vectors" (ISVs) were designed to optimize integration of a yeast-selectable marker specifically into yeast artificial chromosomes (YACs) derived from human but not mouse DNA. ISVs were transformed into a YAC genomic library constructed from DNA of a human-mouse somatic cell hybrid containing chromosome 21 (HSA21) as the only human chromosome. One percent of the yeast in the original library contained HSA21-derived YACs; between 45% and 54% of the yeast recovered after transformation with ISV vectors contained human YACs. Integrative selection provides a rapid means of obtaining a highly enriched population of human chromosome-specific YACs by eliminating the labor-intensive steps of isolating and screening primary transformants. The procedure is biased toward the selection of YACs that contain a large number of targets for homologous recombination; thus, libraries constructed by this procedure will be composed primarily of the largest YACs in the population.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 7788-7791 |
| Number of pages | 4 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Volume | 88 |
| Issue number | 17 |
| State | Published - 1991 |
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Keywords
- Genetic vector
- Homologous recombination
- Yeast artificial chromosome library
ASJC Scopus subject areas
- General
- Genetics
Cite this
Integrative selection of human chromosome-specific yeast artificial chromosomes. / Pavan, William J.; Reeves, Roger H.
In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 88, No. 17, 1991, p. 7788-7791.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Integrative selection of human chromosome-specific yeast artificial chromosomes
AU - Pavan, William J.
AU - Reeves, Roger H
PY - 1991
Y1 - 1991
N2 - Human specific "integrative selection vectors" (ISVs) were designed to optimize integration of a yeast-selectable marker specifically into yeast artificial chromosomes (YACs) derived from human but not mouse DNA. ISVs were transformed into a YAC genomic library constructed from DNA of a human-mouse somatic cell hybrid containing chromosome 21 (HSA21) as the only human chromosome. One percent of the yeast in the original library contained HSA21-derived YACs; between 45% and 54% of the yeast recovered after transformation with ISV vectors contained human YACs. Integrative selection provides a rapid means of obtaining a highly enriched population of human chromosome-specific YACs by eliminating the labor-intensive steps of isolating and screening primary transformants. The procedure is biased toward the selection of YACs that contain a large number of targets for homologous recombination; thus, libraries constructed by this procedure will be composed primarily of the largest YACs in the population.
AB - Human specific "integrative selection vectors" (ISVs) were designed to optimize integration of a yeast-selectable marker specifically into yeast artificial chromosomes (YACs) derived from human but not mouse DNA. ISVs were transformed into a YAC genomic library constructed from DNA of a human-mouse somatic cell hybrid containing chromosome 21 (HSA21) as the only human chromosome. One percent of the yeast in the original library contained HSA21-derived YACs; between 45% and 54% of the yeast recovered after transformation with ISV vectors contained human YACs. Integrative selection provides a rapid means of obtaining a highly enriched population of human chromosome-specific YACs by eliminating the labor-intensive steps of isolating and screening primary transformants. The procedure is biased toward the selection of YACs that contain a large number of targets for homologous recombination; thus, libraries constructed by this procedure will be composed primarily of the largest YACs in the population.
KW - Genetic vector
KW - Homologous recombination
KW - Yeast artificial chromosome library
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UR - http://www.scopus.com/inward/citedby.url?scp=0025743244&partnerID=8YFLogxK
M3 - Article
C2 - 1881915
AN - SCOPUS:0025743244
VL - 88
SP - 7788
EP - 7791
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
SN - 0027-8424
IS - 17
ER -