Difference between revisions of "Part:BBa K4344020"
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<partinfo>BBa_K4344020 short</partinfo> | <partinfo>BBa_K4344020 short</partinfo> | ||
| − | + | UL19 gene, coding for the capsid protein (VP5) of herpes simplex virus with additional His-tag for purification | |
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===Usage and Biology=== | ===Usage and Biology=== | ||
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<partinfo>BBa_K4344020 SequenceAndFeatures</partinfo> | <partinfo>BBa_K4344020 SequenceAndFeatures</partinfo> | ||
| + | The sequence of <i>UL19</i> encoding for HSV-1 major capsid protein was extracted from the complete https://www.ncbi.nlm.nih.gov/nuccore/KF498959.1 human herpesvirus 1 genome. The sequence was human codon optimised using https://www.benchling.com/ Benchling. | ||
| + | The sequence was screened for published antibody binding sites and functional structure elements using the https://www.uniprot.org/ UniProt database. We identified the amino acids 862 to 880 in <i>UL19</i> as a published antibody binding site <span class="scientific-src">(Han <i>et al.</i>, 2019)</span>. Since it is a published antibody site, we anticipate that the corresponding DNA sequence is highly conserved in the HSV genome. Therefore, we chose a sequence for cloning and expression including this structure element. For <i>UL19</i> bases 2446 to 3444 were selected resulting in a fragment of 999 bp in size. The fragment was further modified to mask unwanted restriction sites and ease primer design while keeping the amino acid sequence unchanged. <i>UL19</i> was modified at the 16-18TCC>AGT and 21G>A. A His-tag (5’-CATCACCATCACCATCAC-3’) and a TAA stop codon were added at the 3’ sequence end. | ||
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Latest revision as of 13:19, 12 October 2022
HSV-UL19 AA 816:1148-6xHis
UL19 gene, coding for the capsid protein (VP5) of herpes simplex virus with additional His-tag for purification
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 301
Illegal NgoMIV site found at 448 - 1000COMPATIBLE WITH RFC[1000]
The sequence of UL19 encoding for HSV-1 major capsid protein was extracted from the complete https://www.ncbi.nlm.nih.gov/nuccore/KF498959.1 human herpesvirus 1 genome. The sequence was human codon optimised using https://www.benchling.com/ Benchling. The sequence was screened for published antibody binding sites and functional structure elements using the https://www.uniprot.org/ UniProt database. We identified the amino acids 862 to 880 in UL19 as a published antibody binding site (Han et al., 2019). Since it is a published antibody site, we anticipate that the corresponding DNA sequence is highly conserved in the HSV genome. Therefore, we chose a sequence for cloning and expression including this structure element. For UL19 bases 2446 to 3444 were selected resulting in a fragment of 999 bp in size. The fragment was further modified to mask unwanted restriction sites and ease primer design while keeping the amino acid sequence unchanged. UL19 was modified at the 16-18TCC>AGT and 21G>A. A His-tag (5’-CATCACCATCACCATCAC-3’) and a TAA stop codon were added at the 3’ sequence end.