Nucleic Acids Research, Vol. 18, No. 12 3631
.=) 1990 Oxford University Press
Nucleotide sequence of the HPV16 Li open reading frame Adrian Parton I.C.R.F. Tumour Virus Group, Cambridge CB2 lQP, UK Submitted May 23, 1990
GenBank accession
Double strand plasmid sequencing of an HPV16 LI PCR product revealed two differences from the original published sequence (Seedorf et al., 1985). The Li PCR fragment was derived from genomic DNA using LI 'N' and 'C' terminal specific oligonucleotide primers. The entire open reading frame was sequenced using primers spaced at approximately 200 bp intervals. Two differences from the original published sequence were observed, an insertion of CAT at position 6900 and a deletion of GAT at position 6951. To eliminate the possibility that these differences were not due to PCR artefacts two other DNAs were sequenced, the original pAT153 HPV16 clone (Durst et al., 1983) and W12 DNA (a HPV16 DNA cloned from a cervical keratinocyte cell line (Stanley et al., 1989; Storey, unpublished data). Figure 1 demonstrates that all three sequences were identical and all differ from the original published sequence at nucleotide positions 6900 and 6951. 1 3 2 TCAGTCAGT CAG
no.
K02718
The effects of these nucleotide sequence alterations upon the predicted amino acid sequence are shown in Figure 2. Although the overall open reading frame length remains the same an additional serine residue is inserted between nucleotides 6900 and 6901, which could act as a potential site of phosphorylation.
REFERENCES 1. Durst,M., Gissmann,L., Ikenberg,H. and zur Hausen,J. (1983) Proc. Natl. Acad. Sci. USA 80, 3812-3815. 2. Seedorf,K., Krammer,G., Durst,M., Suhai,S. and Rowenkamp,W. (1985) Virology 145, 181. 3. Stanley,M.A., Browne,H.M., Appleby,M. and Minson,A.C. (1989) Int. J. Cancer 43, 672-676.
Sequence of Li PCR Fragment 6951
CTAAAGAAGATCCCCTTAAAAAA
LYS GLU ASP PRO LEU
-GAT
-----
,' -,;--
s
CGAT
Published HPV 16 Sequence _4MI
AM*;
wx....
6951
_ ...um;
I
~
CTAAAGAAGATGATCCCCTTAAAAAA
LYS GLU ASP ASP PRO LEU
Figure 2. Comparison of the nucleotide sequences and predicted amino acid sequence of the original published sequence (Seedorf et al., 1985) and the LI PCR fragment. The CAT insertion and GAT deletion are shown boxed for identification.
Figure 1. Nucleotide sequences of the LI PCR fragment, (1); the HPV16 pAT153 clone, (2); and W12 DNA, (3). Each track is labelled with the corresponding nucleotide. The positions of the additional CAT sequence and deleted GAT sequence are indicated by arrows.
.=) 1990 Oxford University Press
Nucleotide sequence of the HPV16 Li open reading frame Adrian Parton I.C.R.F. Tumour Virus Group, Cambridge CB2 lQP, UK Submitted May 23, 1990
GenBank accession
Double strand plasmid sequencing of an HPV16 LI PCR product revealed two differences from the original published sequence (Seedorf et al., 1985). The Li PCR fragment was derived from genomic DNA using LI 'N' and 'C' terminal specific oligonucleotide primers. The entire open reading frame was sequenced using primers spaced at approximately 200 bp intervals. Two differences from the original published sequence were observed, an insertion of CAT at position 6900 and a deletion of GAT at position 6951. To eliminate the possibility that these differences were not due to PCR artefacts two other DNAs were sequenced, the original pAT153 HPV16 clone (Durst et al., 1983) and W12 DNA (a HPV16 DNA cloned from a cervical keratinocyte cell line (Stanley et al., 1989; Storey, unpublished data). Figure 1 demonstrates that all three sequences were identical and all differ from the original published sequence at nucleotide positions 6900 and 6951. 1 3 2 TCAGTCAGT CAG
no.
K02718
The effects of these nucleotide sequence alterations upon the predicted amino acid sequence are shown in Figure 2. Although the overall open reading frame length remains the same an additional serine residue is inserted between nucleotides 6900 and 6901, which could act as a potential site of phosphorylation.
REFERENCES 1. Durst,M., Gissmann,L., Ikenberg,H. and zur Hausen,J. (1983) Proc. Natl. Acad. Sci. USA 80, 3812-3815. 2. Seedorf,K., Krammer,G., Durst,M., Suhai,S. and Rowenkamp,W. (1985) Virology 145, 181. 3. Stanley,M.A., Browne,H.M., Appleby,M. and Minson,A.C. (1989) Int. J. Cancer 43, 672-676.
Sequence of Li PCR Fragment 6951
CTAAAGAAGATCCCCTTAAAAAA
LYS GLU ASP PRO LEU
-GAT
-----
,' -,;--
s
CGAT
Published HPV 16 Sequence _4MI
AM*;
wx....
6951
_ ...um;
I
~
CTAAAGAAGATGATCCCCTTAAAAAA
LYS GLU ASP ASP PRO LEU
Figure 2. Comparison of the nucleotide sequences and predicted amino acid sequence of the original published sequence (Seedorf et al., 1985) and the LI PCR fragment. The CAT insertion and GAT deletion are shown boxed for identification.
Figure 1. Nucleotide sequences of the LI PCR fragment, (1); the HPV16 pAT153 clone, (2); and W12 DNA, (3). Each track is labelled with the corresponding nucleotide. The positions of the additional CAT sequence and deleted GAT sequence are indicated by arrows.
