Degenerate PCR method for identification of an antiapoptotic gene in BHV-1
| Autor: | Francesco Napolitano, Maria Gabriella Giganti, Ugo Pagnini, Giuseppe Iovane, Maria Rosa Ciriolo, Gabriella Marfe, L. De Martino, C Di Stefano, P Sinibaldi Salimei, Giuseppe Filomeni |
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| Přispěvatelé: | G., Marfe, DE MARTINO, Luisa, G., Filomeni, C., DI STEFANO, M. G., Giganti, Pagnini, Ugo, F., Napolitano, Iovane, Giuseppe, M. R., Ciriolo, P., SINIBALDI SALIMEI |
| Rok vydání: | 2006 |
| Předmět: |
PCR degenerate
Sequence analysis Amino Acid Motifs Molecular Sequence Data Cell HSP72 Heat-Shock Proteins Apoptosis Biology Polymerase Chain Reaction Biochemistry Cell Line Viral Proteins chemistry.chemical_compound Consensus Sequence medicine Settore MED/05 - Patologia Clinica Animals Humans Settore BIO/10 Molecular Biology Gene DNA Primers Herpesvirus 1 Bovine Genetics Base Sequence Sequence Homology Amino Acid Strain (chemistry) Cell Biology apoptosi Molecular biology CODEHOP medicine.anatomical_structure Databases as Topic chemistry BHV-1 UL14 Sorbitol K562 Cells PCR degenerata Software DNA K562 cells |
| Zdroj: | Journal of Cellular Biochemistry. 97:813-823 |
| ISSN: | 1097-4644 0730-2312 |
| Popis: | To investigate on the hypothetical presence of an antiapoptotic gene, we utilized the CODEHOP (COnsensus-DEgenerate Hybrid Oligonucleotide Primers) strategy amplifying unknown sequences from a background of genomic (bovine herpesvirus type-1) BHV-1 DNA. An alignment of carboxyl-terminal domains belonging to three proteins encoded by g34.5, MyD116 and GADD34 genes, was carried out to design degenerate PCR primers in highly conserved regions. This allowed the amplification of a 110 bp fragment. This fragment was subjected to automatic sequencing and DNA sequence analysis revealed that its position resided between the nt 14363 and the nt 14438 in bovine herpesvirus type-1 (BHV-1) Cooper strain sharing an identity of 86% (UL14). Transient transfections showed that UL14 protein is efficient in protecting MDBK and K562 cells from sorbitol induced apoptosis. The protein's anti-apoptotic func- tion may derive from its heat shock protein-like properties. J. Cell. Biochem. 97: 813-823, 2006. 2005 Wiley-Liss, Inc. |
| Databáze: | OpenAIRE |
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