Alterations in the regulatory region of the human papillomavirus type 6 genome are generated during propagation in Escherichia coli.
AUTOR(ES)
Kasher, M S
RESUMO
We analyzed the long control regions (LCRs) of seven human papillomavirus type 6b (HPV-6b) clones, which contained prototype HPV-6b sequences recloned into various plasmid vectors and propagated in different strains of Escherichia coli. Southern blot analysis and DNA sequencing demonstrated three different sequences, each distinct from the published prototype HPV-6b sequence. Two of the plasmids contained insertions of 24 and 94 base pairs (bp) and a 1-bp deletion. Four plasmids contained insertions of 24 and 58 bp and a deletion of 49 bp. One plasmid contained a single insertion of 77 bp. The 94-, and 58-bp insertions occurred at the same site and had 100% positional identity across their shared lengths. All changes were located in the purine-thymidine-rich region of the LCR (nucleotides 7292 to 7400). Two additional LCR sequences were detected by restriction analysis of two other HPV-6b clones. We conclude that the purine-thymidine-rich region of the LCR is a hot spot for recombination in E. coli and that the alterations are the result of recA-independent events. These results emphasize the need to rigorously prove that a cloned isolate is an authentic copy of the genomic DNA present in the original lesion. In addition, the data indicate that the HPV-6b LCR sequences employed in different laboratories may be different, even if their parental DNAs were identical. Finally, we discuss the need for caution in assigning biological significance to alterations in this region, in view of the limited data available on the true identity of the HPV-6b LCR.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=253450Documentos Relacionados
- Transcriptional regulatory elements in the noncoding region of human papillomavirus type 6.
- Analysis of the regulatory region of the ssb gene of Escherichia coli.
- Two Novel Promoters in the Upstream Regulatory Region of Human Papillomavirus Type 31b Are Negatively Regulated by Epithelial Differentiation
- Genetic Analysis of cis Regulatory Elements within the 5′ Region of the Human Papillomavirus Type 31 Upstream Regulatory Region during Different Stages of the Viral Life Cycle
- Trichostatin A Up-Regulates Human Papillomavirus Type 11 Upstream Regulatory Region-E6 Promoter Activity in Undifferentiated Primary Human Keratinocytes