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Identification of two polymorphic microsatellite repeats on a bovine TNFA-containing BAC clone

Two bovine bacterial artificial chromosome clones were screened by PCR to amplify the tumour necrosis factor alpha (TNFA) gene. DNA extraction and Southern blotting and probing with a (GT)10 oligonucleotide were performed, followed by cloning into the pUC19 vector. A 0.9 kb fragment containing seque...

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Autores principales: Wambugu, J., Kanguha, E., Hanotte, Olivier H., Davis, S., Taylor, J., Skow, L., Brunner, R., Schwerin, M., Teale, A.J., Iraqi, F.A.
Formato: Journal Article
Lenguaje:Inglés
Publicado: 2001
Materias:
cattle
genes
genetic polymorphism
microsatellites
nucleotide sequences
tumour necrosis factor
Acceso en línea:https://hdl.handle.net/10568/28253
Sumario:Two bovine bacterial artificial chromosome clones were screened by PCR to amplify the tumour necrosis factor alpha (TNFA) gene. DNA extraction and Southern blotting and probing with a (GT)10 oligonucleotide were performed, followed by cloning into the pUC19 vector. A 0.9 kb fragment containing sequences homologous with the probe was sequenced and a (GT)9 microsatellite was identified. Further DNA sequencing for the subclones yielded a (TA)10(TTA)5 microsatellite. Primers were designed to amplify these 2 microsatellites in 4 cattle breeds. The PCR product length was polymorphic as shown by denaturing polyacrylamide gel electrophoresis. The microsatellites were designated as ILFI-TNF-I and II for (GT)9 and (TA)10(TTA)5, respectively.

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