Further Annotation of Phage Elesar Genes 12 and 13

Date: 2-4-19

• Rational
  • The rational for this lab was to annotate the function data for two genes from phage Elesar in order to gain practice in annotation before transitioning to phage NapoleonB.
• Procedure
  1. DNA Master was opened.
  2. File > Open > Archived DNA Master File was used to open the same Elesar FastA file from before.
  3.  Gene 12 was selected through the listing of predicted genes, and the Product was copied.
  4. The amino acid sequence was loaded into the BLASTp feature on the NCBI website and the BLAST results were analyzed.
  5. The amino acid sequence was loaded into the BLAST feature on the PhagesDB website and the BLAST results were analyzed.
  6.  Gene 13 was selected through the listing of predicted genes, and the Product was copied.
  7. The amino acid sequence was loaded into the BLASTp feature on the NCBI website and the BLAST results were analyzed.
  8. The amino acid sequence was loaded into the BLAST feature on the PhagesDB website and the BLAST results were analyzed.
  9. Using the acquired information, the 12th and 13th genes were annotated.
• Results
  • The following annotations were made for Elesar gene 12 :
    • F: The function was determined to be a head-to-tail stopper. This was supported by the following fields.
    • SIF-BLAST: head-to-tail stopper, NCBI BLAST, Arthrobacter Phage Nandita gp13, AYN58635.1, 87%, 5e-58
    • SIF-HHPred: NKF, Pfam-A, DUF3599, PF12206.8, 100%, 99.09
    • SIF-Syn: head-to-tail stopper, upstream gene is head-to-tail adaptor like in arthrobacter phage Ryan, downstream gene has NKF
  • The following annotations were made for Elesar gene 13 :
    • F: NKF. This was deternined because there no significant blast hits pointing to a function.
    • SIF-BLAST: NKF
    • SIF-HHPred: NKF
    • SIF-Syn: NKF
• Conclusion
  • Phage Elesar gene 12 has a probably function of being a head-to-tail stopper due to its similarities to other head-to-tail stopper genes as well as its synteny.
• Future Plans
  • The next step is to continue annotating more genes on Elesar for practice before transitioning to NapoleonB.