PMID- 9371404
OWN - NLM
STAT- MEDLINE
DCOM- 19971209
LR  - 20180214
IS  - 0301-0171 (Print)
IS  - 0301-0171 (Linking)
VI  - 78
IP  - 2
DP  - 1997
TI  - Generation of chicken Z-chromosome painting probes by microdissection for 
      screening large-insert genomic libraries.
PG  - 124-30
AB  - A strategy for rapid generation of chicken sex chromosome-Z painting probes has 
      been developed using microdissection. Whole chromosome painting probes (WCPs) 
      were prepared from 10-15 copies of mitotic metaphase chicken Z chromosomes. The 
      microisolated chromosomes were subjected to PEG/proteinase K treatment in a 
      collection drop to release DNA, which was then amplified using a degenerate 
      oligonucleotide-primed shuttle PCR (DOP-Shuttle-PCR) strategy. Size distributions 
      of the PCR products were analyzed by agarose gel electrophoresis and smears of 
      DNA were revealed that ranged in size from 200-800 bp, without any evidence of 
      preferential amplification. Both specificity and complexity of the probes have 
      been analyzed by Southern blot and fluorescence in situ hybridization (FISH). 
      Non-specific hybridization was efficiently blocked by using chicken competitor 
      DNA. Analysis of the WCPs produced shows that collectively they provide uniform 
      hybridization signals along the entire length of the chicken Z chromosome. To 
      demonstrate one possible application of these complex probes, we screened a 
      large-insert bacterial artificial chromosome (BAC) chicken genomic library to 
      select Z chromosome-specific clones. To address specificity of the selected 
      clones and to physically map them to the Z chromosome, FISH analysis was used. Of 
      the 3 clones initially tested, one clone (C3) carrying a 250-kb insert mapped to 
      the distal portion of the short arm of the chicken Z chromosome. Therefore, this 
      technique has provided appropriate probes for screening large-insert genomic 
      libraries. Further application of these probes includes the analysis of 
      chromosome rearrangements, studies of cases of heteroploidy involving the Z 
      chromosome, positional cloning of Z-linked genes and studies on mechanisms of 
      sex-chromosome evolution in birds.
FAU - Zimmer, R
AU  - Zimmer R
AD  - Department of Animal and Poultry Science, University of Guelph, Ontario, Canada.
FAU - King, W A
AU  - King WA
FAU - Verrinder Gibbins, A M
AU  - Verrinder Gibbins AM
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Switzerland
TA  - Cytogenet Cell Genet
JT  - Cytogenetics and cell genetics
JID - 0367735
SB  - IM
MH  - Animals
MH  - Cells, Cultured
MH  - Chickens/*genetics
MH  - *Chromosome Mapping
MH  - *Chromosomes
MH  - *Gene Library
MH  - Polymerase Chain Reaction/methods
EDAT- 1997/01/01 00:00
MHDA- 1997/11/26 00:01
CRDT- 1997/01/01 00:00
PHST- 1997/01/01 00:00 [pubmed]
PHST- 1997/11/26 00:01 [medline]
PHST- 1997/01/01 00:00 [entrez]
AID - 10.1159/000134643 [doi]
PST - ppublish
SO  - Cytogenet Cell Genet. 1997;78(2):124-30. doi: 10.1159/000134643.