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.
Publication Date: 1998-07-29 PubMed ID: 9680383DOI: 10.1007/s003359900835Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article discusses the creation of a horse Bacterial Artificial Chromosome (BAC) library and its use in identifying and locating specific sequences within horse chromosomes. The library, which is a collection of cloned DNA fragments, aimed to broaden understanding of equine genetics and potentially aid in finding genes of interest.
Construction of the BAC library
- The researchers created a horse BAC library consisting of approximately 40,000 clones, with an average insert size of 110 kilobases (kb). This represented 1.5 times the coverage of the horse genome.
- Having multiple copies of the genome increases the chances of finding a particular DNA sequence. The researchers assessed that for this BAC library, the likelihood of finding a specific sequence was 0.75.
Characterization and Localization of the BAC library
- The team characterized the BAC library by using Polymerase Chain Reaction (PCR) to screen around 130 sequences of horse microsatellites and exonic gene sequences obtained from various databases.
- Following characterization, particular BAC clones containing 8 microsatellites and 12 genes were isolated and precisely placed in the horse’s chromosomes, a process known as fluorescent in situ hybridization (FISH).
Assigning and Orienting Linkage Groups
- The researchers assigned two linkage groups (groups of genes that tend to be inherited together) — LG2 and LG5 — to specific horse chromosomes: ECA3 and ECA24 respectively.
- Further, they managed to orient (determine the relative positions of) another five linkage groups: LG3, LG4, LG5, LG8, and LG12. Only three groups remained unassigned.
Significance of the BAC library
- The construction of this BAC library is a significant step towards creating an integrated map of the entire horse genome, which could become a reality in the near future.
- Such a comprehensive genetic map has the potential to improve comparative mapping processes, helping scientists to more efficiently locate and investigate candidate genes of interest – genes that may be linked to specific traits or diseases in horses.
Cite This Article
APA
Godard S, Schibler L, Oustry A, Cribiu EP, Guérin G.
(1998).
Construction of a horse BAC library and cytogenetical assignment of 20 type I and type II markers.
Mamm Genome, 9(8), 633-637.
https://doi.org/10.1007/s003359900835 Publication
Researcher Affiliations
- INRA, Centre de Recherche de Jouy, Laboratoire de Génétique biochimique et de Cytogénétique, 78352 Jouy-en-Josas, Cedex, France.
MeSH Terms
- Animals
- Chromosome Mapping
- DNA Primers
- Exons
- Gene Library
- Genetic Markers
- Horses / genetics
- In Situ Hybridization, Fluorescence
- Microsatellite Repeats
- Polymerase Chain Reaction
Citations
This article has been cited 9 times.- Liu C, Bai C, Guo Y, Liu D, Lu T, Li X, Ma J, Ma Y, Guan W. Construction and analysis of Siberian tiger bacterial artificial chromosome library with approximately 6.5-fold genome equivalent coverage.. Int J Mol Sci 2014 Mar 7;15(3):4189-200.
- Liu C, Guo Y, Lu T, Wu H, Na R, Li X, Guan W, Ma Y. Construction and preliminary characterization analysis of Wuzhishan miniature pig bacterial artificial chromosome library with approximately 8-fold genome equivalent coverage.. Biomed Res Int 2013;2013:587493.
- Wallner B, Vogl C, Shukla P, Burgstaller JP, Druml T, Brem G. Identification of genetic variation on the horse y chromosome and the tracing of male founder lineages in modern breeds.. PLoS One 2013;8(4):e60015.
- Hamilton NA, Tammen I, Raadsma HW. Multi-species comparative analysis of the equine ACE gene identifies a highly conserved potential transcription factor binding site in intron 16.. PLoS One 2013;8(2):e55434.
- Chowdhary BP, Raudsepp T. The horse genome derby: racing from map to whole genome sequence.. Chromosome Res 2008;16(1):109-27.
- Lindgren G, Breen M, Godard S, Bowling A, Murray J, Scavone M, Skow L, Sandberg K, Guérin G, Binns M, Ellegren H. Mapping of 13 horse genes by fluorescence in-situ hybridization (FISH) and somatic cell hybrid analysis.. Chromosome Res 2001;9(1):53-9.
- Shiue Y-L, Millon LV, Skow LC, Honeycutt D, Murray JD, Bowling AT. Synteny and regional marker order assignment of 26 type I and microsatellite markers to the horse X- and Y-chromosomes.. Chromosome Res 2000;8(1):45-55.
- Caetano AR, Shiue YL, Lyons LA, O'Brien SJ, Laughlin TF, Bowling AT, Murray JD. A comparative gene map of the horse (Equus caballus).. Genome Res 1999 Dec;9(12):1239-49.
- Lindgren G, Sandberg K, Persson H, Marklund S, Breen M, Sandgren B, Carlstén J, Ellegren H. A primary male autosomal linkage map of the horse genome.. Genome Res 1998 Sep;8(9):951-66.
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