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All posts created by debbie

| posted 25 Feb, 2019 23:00
Instead , its function can be denoted as “HTH DNA binding protein, MerR-like”.

The folks at LeTourneau University did not agree with the funcitonal assignemnt of "peptidase" to Glaske gp65 (coordinates:43072 to 43974).

According to Dr. Frederick, Fred Ballraine and students did some investigating.
"Thirty one of the 103 hits above 90% probability reference “MerR”. We find nothing in the HHPred results referencing a “peptidase” function or domain. However, there are five references to peptidase function in the NCBI Blast results. All are phages. While I did not dig deeper, most are ours. The first hit “Babsiella” lists the two of you, Dan and Graham as authors on the GenBank submission (201smile. So I suspect we are missing some evidence you all saw.

A local BlastP on PhagesDB with Glaske gp65 hits 16 of our SEA phages identifying the function of this proteins as a “peptidase”.

Since HHPred does not reference “peptidase” once for any of the hits and since an NCBI BlastP does not reference any other than our phages and a few other phages, I suggest that someone jumped at the “peptidase” function for this gene. Then once it was in the database and on the “Official Function List” everyone started to “Band-wagon” as one of my student’s likes to put it.

We are going to list the function as “HTH DNA binding protein, MerR-like”.
Edited 25 Feb, 2019 23:02
Posted in: Cluster P Annotation TipsGlaske gp65 is not a peptidase
| posted 25 Feb, 2019 22:38
Some Cluster EA phages have a tRNA.
Right now (2-14-19); it looks like the the Clusters EA3 and EA4 have one.
Posted in: Cluster EA Annotation TipstRNA in Cluster EA
| posted 25 Feb, 2019 22:26
Susan,
Excellent question! I think those tRNAs are missed in Golden and Koji. I will be making some edits soon.
Good catch!
debbie
Posted in: tRNAsEA1 tRNA
| posted 20 Feb, 2019 17:05
Hi all,
I can weigh in on this. In the papers that we wrote about Cluster O, J, and M, we did some mass spec. The data showed that where there were 2 starts in a row, no methionine was present in that mass spec data. The interpretation of that is likely that post-translational chopping of the initiation methionine occurred. (So there could only be one present to chop.)
I just scanned the papers to see if we wrote that in one of them but couldn't find it. So by all means take a look to see if it is there.
But that is why we suggest that the second start codon (1 base overlap) be called. It is hard to do, when you are progammed (like me) to look for the 4-base pair overlap and all other data - usually the SD score - points to the first start codon as the better call. But you can do it! It also was not done on lots of files and it is difficult to pinpoint to fix.
Posted in: Choosing Start SitesGTGA overlaps
| posted 11 Feb, 2019 22:12
I believe that GeneMark is now working!
Back in business!
debbie
Posted in: DNA MasterAuto-annotation fix for fall 2017 and later
| posted 11 Feb, 2019 11:20
Dana,
Hi. I believe that the issue is a licensing issue that we need to do on our end. It should be fixed later today.
debbie
Posted in: DNA MasterAuto-annotation fix for fall 2017 and later
| posted 05 Feb, 2019 20:00
Rick,
You are right, because Emporer and the prophage in G. westfalica are identical sequences, the Emporer hit is buried. Check out the alignment, you will see +2 more titles. You will find Emporer there.
Posted in: AnnotationPhage hits not showing in NCBI but 100% to Gordonia wetsfalica
| posted 05 Feb, 2019 17:53
Rick,
I believe the simple explanation is that the BLASTp database(s) have not incorporated the Emperor submission yet. Which, I agree seems odd.
Posted in: AnnotationPhage hits not showing in NCBI but 100% to Gordonia wetsfalica
| posted 01 Feb, 2019 03:50
Hi Denise,
If you find one transmembrane domain with TMHMM, you can only call it a membrane protein if you can validate it with a different transmembrane finder (such as SOSUI). (If two different algorithms detect a trans membrane domain, you can hopefully rule out false positives.)
Does that clarify it?
debbie
Posted in: Functional AnnotationMembrane protein
| posted 22 Jan, 2019 01:54
The infernal score for the tRNAs called is lower than what is described in the guide. Use an infernal score of 20 for the tRNA calls in Cluster C annotations.
Posted in: Cluster C Annotation TipstRNAs in Cluster C