Rationale: In lab today, feedback surrounding annotations from Wednesday was given. Therefore, it was found to be pertinent to correct mistakes that had been made, and additional processing techniques, such as HHPred, were learned and utilized.

Tools:

• DNA Master
• HHPred
• NCBI BLAST
• PhagesDB BLAST

Procedure:

• Genes 50 and 51 of Elesar were re-annotated, with the LO and RBS categories being fixed. SIF-HHPred and SIF-BLAST was also ran for both genes.

Results:

Photos show results of gene 50 and 51, respectively.

Gene 50:

SSC: 36095,36970 CP: Yes SCS: Both BLAST-Start: Nandita, gp50, NCBI, Query 1 to Subject 1, 99%, 0.0. Gap: 40. LO: No RBS: Kibler7,Kibler Medium, 2.097, -4.366, Yes. SIF- BLAST: DNA Polymerase, Nandita gp50, AYN58673.1, 96%, 0.0 SIF- HHPred: DNA Polymerase III Beta, pfam, PF02767.16, 99.48, 5.8e-16

Gene 51:

SSC: 36967,37350 CP: Yes SCS: Both. BLAST-Start: Auxilium, gp 51, NCBI, Query 1 to Subject 1, 96%, 4e-14. Gap: 4bp overlap. LO: N/A RBS: Kibler7, Kibler Medium, 2.496, -3.662, Yes. SIF- BLAST: NKF. SIF- HHPred: NKF.

Conclusions: Since all evidence for the first gene points to one specific function, it can be called that the function of gene 50 of Elesar is to code for DNA Polymerase III. No such call can be made for gene 51, as only a hypothetical protein was called for this sequence, with no substantial support from the SIF-HHPred.

Next Steps:

The next step that will happen is that skills that have been developed through the past few labs will begin to be applied to NapoleonB. The only other skill that needs to be developed is the use of starterator, which will occur at a later date.