TY - JOUR
T1 - Physical mapping of plasmid and cosmid clones in filamentous fungi by fiber-FISH
AU - Tsuchiya, Dai
AU - Matsumoto, Aya
AU - Covert, Sarah F.
AU - Bronson, Charlotte R.
AU - Taga, Masatoki
N1 - Funding Information:
We thank Olen C. Yoder and B. Gillian Turgeon for providing plasmids p627PKSCH#28 (PKS1) and pC4DEC1 (DEC1) as well as helpful suggestions. Thanks are also to Hans D. VanEtten for supplying strain 156-30-6. This work was supported in part by Grant-in-Aid No. 13660053 from Japan Society for the Promotion of Science to M. Taga.
PY - 2002
Y1 - 2002
N2 - Fluorescence in situ hybridization to extended DNA fibers (fiber-FISH) serves as a powerful tool for direct physical mapping in plants and animals. Here, we show that fiber-FISH is useful for contig mapping as well as for estimating the physical distance between genetic markers in fungi. A five-cosmid contig from a chromosome of Nectria haematococca and four cloned genetic markers from a linkage map of Cochliobolus heterostrophus were chosen as models for the application of this technology. In N. haematococca, overlapping and non-overlapping clones were visually mapped on individual DNA fibers, confirming the results from conventional physical mapping perfectly. Fiber-FISH concomitantly indicated the gap size or the extent of overlap between two clones. In C. heterostrophus, the physical distance between the two pairs of genetic markers could be estimated from the microscopic measurements of the intervals. Chromosomal DNA isolated from a pulsed field gel was suitable for preparing the DNA fibers.
AB - Fluorescence in situ hybridization to extended DNA fibers (fiber-FISH) serves as a powerful tool for direct physical mapping in plants and animals. Here, we show that fiber-FISH is useful for contig mapping as well as for estimating the physical distance between genetic markers in fungi. A five-cosmid contig from a chromosome of Nectria haematococca and four cloned genetic markers from a linkage map of Cochliobolus heterostrophus were chosen as models for the application of this technology. In N. haematococca, overlapping and non-overlapping clones were visually mapped on individual DNA fibers, confirming the results from conventional physical mapping perfectly. Fiber-FISH concomitantly indicated the gap size or the extent of overlap between two clones. In C. heterostrophus, the physical distance between the two pairs of genetic markers could be estimated from the microscopic measurements of the intervals. Chromosomal DNA isolated from a pulsed field gel was suitable for preparing the DNA fibers.
KW - Chromosome
KW - Cochliobolus heterostrophus
KW - Contig
KW - Fiber-FISH
KW - Fluorescence in situ hybridization
KW - Nectria haematococca
KW - Physical mapping
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U2 - 10.1016/S1087-1845(02)00026-9
DO - 10.1016/S1087-1845(02)00026-9
M3 - Article
C2 - 12223186
AN - SCOPUS:0036387167
VL - 37
SP - 22
EP - 28
JO - Fungal Genetics and Biology
JF - Fungal Genetics and Biology
SN - 1087-1845
IS - 1
ER -