The application of AFLP fingerprinting to construct a YAC contig containing ADH2 and MTP on sheep chromosome 6.
The low density of genetic markers on livestock maps limits progress in positional cloning projects. We demonstrate a strategy of combining comparative mapping with AFLP fingerprinting to develop physical maps in a defined region of the sheep genome. Sequence tagged sites for alcohol dehydrogenase 2 (ADH2) and microsomal triglyceride transfer protein (MTP) were developed and used to screen a sheep yeast artificial chromosome (YAC) library. Nine YACs were identified containing the microsatellite marker BM1329 and either ADH2 or MTP. Additional markers in the region were not available, and AFLP analysis was developed to identify sheep-specific bands within the YACs to determine their degree of overlap. Fourteen bands common to more than one YAC were analysed and provided the markers necessary to develop a YAC contig containing the three STS markers. One YAC (yac260B5) containing all three markers (ADH2, MTP, and BM1329) was mapped to sheep chromosome 6q1.6-->q1.8 by FISH analysis.
AgResearch Molecular Biology Unit and the Centre for Gene Research, Department of Biochemistry, University of Otago, Dunedin, New Zealand.
J M
JM
Lumsden
E A
EA
Lord
S A
SA
Cato
T E
TE
Richardson
T C
TC
van Stijn
M F
MF
Broom
K
K
Patel
G W
GW
Montgomery
D000426:Alcohol Dehydrogenase / Q000235:genetics*; D000818:Animals; D002352:Carrier Proteins / Q000235:genetics*; D002875:Chromosomes / Q000235:genetics*; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics; D020451:Contig Mapping / Q000379:methods*; D005819:Genetic Markers / Q000235:genetics; D015698:Genomic Library; D017404:In Situ Hybridization, Fluorescence; D018895:Microsatellite Repeats / Q000235:genetics; D012150:Polymorphism, Restriction Fragment Length; D016324:Sequence Tagged Sites; D012756:Sheep / Q000235:genetics*
D002352:Carrier Proteins; D005819:Genetic Markers; C106092:microsomal triglyceride transfer protein; D000426:Alcohol Dehydrogenase