Gene identification and analysis of transcripts differentially regulated in fracture healing by EST sequencing in the domestic sheep.
BACKGROUND
The sheep is an important model animal for testing novel fracture treatments and other medical applications. Despite these medical uses and the well known economic and cultural importance of the sheep, relatively little research has been performed into sheep genetics, and DNA sequences are available for only a small number of sheep genes.


RESULTS
In this work we have sequenced over 47 thousand expressed sequence tags (ESTs) from libraries developed from healing bone in a sheep model of fracture healing. These ESTs were clustered with the previously available 10 thousand sheep ESTs to a total of 19087 contigs with an average length of 603 nucleotides. We used the newly identified sequences to develop RT-PCR assays for 78 sheep genes and measured differential expression during the course of fracture healing between days 7 and 42 postfracture. All genes showed significant shifts at one or more time points. 23 of the genes were differentially expressed between postfracture days 7 and 10, which could reflect an important role for these genes for the initiation of osteogenesis.


CONCLUSIONS
The sequences we have identified in this work are a valuable resource for future studies on musculoskeletal healing and regeneration using sheep and represent an important head-start for genomic sequencing projects for Ovis aries, with partial or complete sequences being made available for over 5,800 previously unsequenced sheep genes.
Hecht
Jochen
J
Max Planck Institute for Molecular Genetics, Berlin, Germany. hecht@molgen.mpg.de
Kuhl
Heiner
H
Haas
Stefan A
SA
Bauer
Sebastian
S
Poustka
Albert J
AJ
Lienau
Jasmin
J
Schell
Hanna
H
Stiege
Asita C
AC
Seitz
Volkhard
V
Reinhardt
Richard
R
Duda
Georg N
GN
Mundlos
Stefan
S
Robinson
Peter N
PN
D000818:Animals; D016000:Cluster Analysis; D020451:Contig Mapping / Q000379:methods; D020224:Expressed Sequence Tags; D017102:Fracture Healing / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods*; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D017422:Sequence Analysis, DNA / Q000379:methods; D034561:Sheep, Domestic / Q000235:genetics*