Expression screening and annotation of a zebrafish myoblast cDNA library.
To analyse the myogenic transcriptome and identify novel genes involved in muscle development in an in vivo context, we have constructed a muscle specific cDNA library from GFP-expressing myoblasts purified by fluorescent activated cell sorting of transgenic zebrafish embryos. We have generated 153,428 EST sequences from this library that have been clustered into consensi, mapped to the genome assembly Zv6 and analysed for protein homology. Expression analysis of a randomly picked sample of clones using whole mount in situ hybridisation, identified 30 genes that are expressed specifically within the myotome, one third of which represent novel sequences. These genes have been assigned to syn-expression groups. The sequencing of the myoblast enriched cDNA library has significantly increased the number of zebrafish ESTs, facilitating the prediction of new spliced transcripts in the genome assembly and providing a transcriptome of an in vivo myoblast cell.
Baxendale
Sarah
S
MRC Centre for Developmental and Biomedical Genetics, University of Sheffield, Sheffield S10 2TN, UK.
Chen
Chao-Kung
CK
Tang
Haizhou
H
Davison
Claire
C
Hateren
Lisa Van
LV
Croning
Mike D R
MD
Humphray
Sean J
SJ
Hubbard
Simon J
SJ
Ingham
Philip W
PW
D000818:Animals; D030801:Animals, Genetically Modified; D004625:Embryo, Nonmammalian; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling / Q000379:methods*; D015723:Gene Library; D023281:Genomics / Q000379:methods; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D032446:Myoblasts / Q000378:metabolism*; D009928:Organ Specificity / Q000235:genetics; D017422:Sequence Analysis, DNA / Q000379:methods*; D015027:Zebrafish / Q000196:embryology / Q000235:genetics* / Q000378:metabolism
D049452:Green Fluorescent Proteins