PMID- 8908520
OWN - NLM
STAT- MEDLINE
DCOM- 19970212
LR  - 20190606
IS  - 1088-9051 (Print)
IS  - 1088-9051 (Linking)
VI  - 6
IP  - 10
DP  - 1996 Oct
TI  - Utilization of FISH in positional cloning: an example on 13q22.
PG  - 1002-12
AB  - In positional cloning the initial assignment of a gene to a specific chromosomal 
      locus is followed by physical mapping of the critical region. The construction of 
      a high-resolution physical map still involves considerable effort. However, new 
      high-resolution fluorescence in situ hybridization (FISH) techniques have 
      facilitated this process substantially. Here we summarize a strategy that 
      combines a spectrum of FISH techniques [metaphase, interphase, mechanically 
      stretched chromosomes (MSCs), and fiber-FISH on free chromatin] for the 
      construction and characterization of a high-resolution physical map for a 
      positional cloning project. The chromosomal region 13q22, containing the locus of 
      the variant form of the neuronal ceroid lipofuscinosis (vLINCL, CLN5) disease, 
      serves here as an example for this process. We used metaphase FISH to exclude 
      positionally a candidate gene, to refine the locus to 13q22, and to analyze the 
      possible chimerism of the YACs in the region. Both metaphase and interphase FISH 
      techniques were applied to determine the low-resolution distances between the 
      restricting markers. FISH using MSCs confirmed the centromeric-telomeric order of 
      the clones and facilitated the estimation of the size of the gaps between the 
      clones. Finally, fiber-FISH was found to be the method of choice for the 
      construction of an accurate high-resolution map of the contig established over 
      the restricted region. Thus, FISH techniques in combination with genetic mapping 
      data enabled the refinement of the initial 4-cM region to a high-resolution map 
      of only 400 kb in length. Here the FISH strategy replaced the need for many 
      laborious traditional physical mapping methods, e.g., pulsed-field gel 
      electrophoresis.
FAU - Laan, M
AU  - Laan M
AD  - Department of Clinical Chemistry, University of Helsinki, Finland.
FAU - Isosomppi, J
AU  - Isosomppi J
FAU - Klockars, T
AU  - Klockars T
FAU - Peltonen, L
AU  - Peltonen L
FAU - Palotie, A
AU  - Palotie A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Genome Res
JT  - Genome research
JID - 9518021
RN  - 0 (Genetic Markers)
RN  - EC 3.4.11.- (Aminopeptidases)
RN  - EC 3.4.14.- (Dipeptidyl-Peptidases and Tripeptidyl-Peptidases)
RN  - EC 3.4.14.10 (tripeptidyl-peptidase 2)
RN  - EC 3.4.21.- (Serine Endopeptidases)
SB  - IM
MH  - Aminopeptidases
MH  - Chromosome Mapping
MH  - Chromosomes, Artificial, Yeast
MH  - *Chromosomes, Human, Pair 13
MH  - *Cloning, Molecular
MH  - Cosmids
MH  - Dipeptidyl-Peptidases and Tripeptidyl-Peptidases
MH  - Genetic Linkage
MH  - Genetic Markers
MH  - Humans
MH  - *In Situ Hybridization, Fluorescence
MH  - Neuronal Ceroid-Lipofuscinoses/genetics
MH  - Polymorphism, Genetic
MH  - Serine Endopeptidases/genetics
EDAT- 1996/10/01 00:00
MHDA- 1996/10/01 00:01
CRDT- 1996/10/01 00:00
PHST- 1996/10/01 00:00 [pubmed]
PHST- 1996/10/01 00:01 [medline]
PHST- 1996/10/01 00:00 [entrez]
AID - 10.1101/gr.6.10.1002 [doi]
PST - ppublish
SO  - Genome Res. 1996 Oct;6(10):1002-12. doi: 10.1101/gr.6.10.1002.