Construction of a horse BAC library and cytogenetical assignment of 20 type I and type II markers

Abstract

A horse BAC library was constructed with about 40,000 clones and mean insert size of 110 kb representing a 1.5 genome equivalent coverage and a probability of finding a single sequence of 0.75. It was characterized by PCR screening of about 130 sequences of horse microsatellites and exonic gene sequences retrieved from databases. BACs containing 8 microsatellites and 12 genes were subsequently localized by fluorescent in situ hybridization (FISH) on chromosomes. Two linkage groups were newly assigned to chromosomes: LG2 to ECA3 and LG5 to ECA24, and five linkage groups were also oriented--LG3, LG4, LG5, LG8, and LG12--leaving only three groups unassigned. This work showed how this library makes an integrated map a realistic objective for the near future and how it can make comparative mapping more efficient in a search for candidate genes of interest.

A specific pattern of splicing for the horse ?S1-Casein mRNA and partial genomic characterization of the relevant locus

“…Clones were prepared as described previously [7,26] and their gene content was ascertained by PCR performed on miniprepped BAC DNA with the same screening primers. PCR fragments were purified with the QIAquick PCR purification kit (Qiagen) and sequenced on an ABI 377 automated sequencer (Applied Biosystems Inc.).…”

“…The BAC insert size was estimated after NotI digestion followed by field inverted gel electrophoresis (FIGE). BAC clones containing the casein locus were used for fluorescence in situ hybridization (FISH) mapping as described elsewhere [1,7].…”

“…Equine genomic DNA was prepared from peripheral blood, as previously described [7]. The Expand TM long template PCR system of BoehringerMannheim (Meylan, France) was used according to the manufacturer's protocol except that annealing was performed at 55…”

Construction and preliminary characterization of a river buffalo bacterial artificial chromosome library

“…BAC libraries have been constructed for several species of agricultural importance, including cattle (Cai et al, 1995;Zhu et al, 1999;Buitkamp et al, 2000;Warren et al, 2000;Eggen et al, 2001;Fujisaki et al, 2002), goat , horse (Godard et al, 1998), pig (Rogel-Gaillard et al, 1999Anderson et al, 2000;Suzuki et al, 2000;Fahrenkrug et al, 2001;Jeon et al, 2003;Liu et al, 2010), and sheep (Gill et al, 1999;Vaiman et al, 1999). Although large-insert DNA libraries from closely related ruminant species such as cattle, goat, and sheep have been used for in situ hybridization experiments in buffalo (Di Meo et al, 2005Perucatti et al, 2009), a buffalo library is necessary for whole-genome physical mapping and isolation of genes and gene clusters as well as for the identification of regulatory elements.…”

Equine protease inhibitor system as a marker for the diagnosis of chronic obstructive pulmonary disease (COPD)

“…Identification of Spi variants using electrophoresis and the measurements of serum Spi concentration by ELISA are not considered ideal procedures (Dagleish et al, 1999(Dagleish et al, , 2000, and the molecular sequence of each Spi has still not been fully determined. It therefore appears that there is a need to establish more efficient methods to identify the genetic variants derived from each Spi (Godard et al, 1998;Milenkovic et al, 2002) in order to clarify the role of this locus in the onset of equine COPD.…”