SEA-PHAGES Logo

The official website of the HHMI Science Education Alliance-Phage Hunters Advancing Genomics and Evolutionary Science program.

Welcome to the forums at seaphages.org. Please feel free to ask any questions related to the SEA-PHAGES program. Any logged-in user may post new topics and reply to existing topics. If you'd like to see a new forum created, please contact us using our form or email us at info@seaphages.org.

All posts created by debbie

| posted 29 Jul, 2022 15:32
Hi Kristen,
As of July 29, 2022, 11:30 am, auto-annotation is back up.
debbie
Posted in: DNA MasterGlimmer failure during autoannotation
| posted 18 Jul, 2022 23:43
Fernando,
Yes, the immunity repressor is called correctly in those genomes. And yes, that area is omitted in Caviar, so you most likely have a lytic phage. Likely a derivative a temperant one.
debbie
Posted in: Cluster A Annotation TipsCluster A3 Immunity Repressor?
| posted 14 Jul, 2022 19:22
Hi Amanda,
Here is thought, that may help. You can BLAST a subset of genes at a time in DNA Master. Here is the Bioinformatics Guide entry that explains:
https://seaphagesbioinformatics.helpdocsonline.com/article-70
Posted in: DNA MasterDNAMaster BLAST failure
| posted 07 Jul, 2022 23:46
Have you wondered what the settings for HHPred, NCBI-blastp, and CDD are used by PECAAN?
The parameters that we run for HHPred are as follows:
Hhsuitedb: "mmcif70/pdb70 NCBI_CD/NCBI_CD scope70/scope70 pfama/pfama",
Proteomes: "",
MsaGenMethod: "UniRef30",
MsaGenMaxIter: "3",
HhpredInclEval: "1e-3",
MinSeqidQuery: "0",
MinCov: "20",
SSScoring: "2",
AlignMacMode: "loc",
MacThreshold: "0.3",
Desc: "250",
Pmin: "20",

and
Blastp from ncbi blast 2.13.0+ is ran remotely with
-db nr
-gapopen 11
-gapextend 1
-word_size 6
-threshold 21
-window_size 40
-evalue 0.05
-max_target_seqs 100
-outfmt 5
-remote
and
The CDD database is searched by rpsblast from ncbi blast 2.13.0+ with the following arguments

-num_alignments=50
-evalue 0.01

7/7/2022
Edited 07 Jul, 2022 23:48
Posted in: PECAANHHPred, Blast, and CDD Parameters
| posted 05 Jul, 2022 18:26
June 21, 2022
Rick Pollenz and Simon White have confirmed (experimentally) that Akoni_gp33 (Cluster EK) is the major capsid subunit! Please annotate! The homolog in Cluster EM is Burro_gp29.
Posted in: Cluster EM Annotation TipsMajor Capsid Protein
| posted 17 Jun, 2022 13:31
Hi all,
As of today, June 17, 2022, there are 88 Cluster EE genomes in our records. They are closely related genomes that contain only ~28 genes. In an effort to make the records congruent, my students and I have reviewed ~75 of them and revised 71 of the records. The template we used is attached here, along with a list of 71 records that we touched. We spent an abundant amount of time on the helix-turn-helix DNA binding proteins at the right end of the genome. We investigated 'types' of helix-turn-helix choices and decided the best path is to call them helix-turn-helix DNA binding proteins.
August 23, 2022 - I just received word that the genomes that we sent to GenBank have been processed. This cluster just might have congruent calls!
Edited 23 Aug, 2022 16:13
Posted in: Cluster EE Annotation TipsGenome Curation - a must read!
| posted 16 Jun, 2022 18:05
Kathleen and Chris,
After reading Chris' entry, he is right -the slippage called in Satis, though it looks very slippery, is not canonical, i.e. not found in the literature. To that end, if we indeed are not calling any of the slippages, I would be inclined to NOT assign the function of a tail assembly chaperone (because there is no HHPred evidence of a TAC) unless I overlooked something.
Posted in: Cluster BM Annotation TipsFrameshift in BM cluster phages?
| posted 16 Jun, 2022 17:52
I just looked at the tail assembly chaperones of Cluster BM.
Here is my take:
currently there are six members:
Satis
Kradla
EhyElimyoE
JustBecause
Kela
Frankenweenie

To the best of my knowledge none of the protein sequences in question here show an HHPred hit to a tail assembly chaperone (TAC). In order to call them TACs, if they possessed a canonical slippage, they could be called TACs and the slippage would be annotated.

The ribosomal slippage found in Satis looks great, AND is identical to what is in Kradal and EhyElimyoE (though they are not called….. yet).

Neither JustBecause or Kela have that same slippage.

And Frankenweenie doesn't look at bit like the others in the c-terminus. (do a blastp - sequence alignment with Satis_58 and you will see how the alignment falls apart.

However, the sequence homology with Satis is compelling. i would be inclined to call both genes TACs, but do not slip them together.

I think that is what I would do.
Good luck!
debbie
Posted in: Cluster BM Annotation TipsFrameshift in BM cluster phages?
| posted 16 Jun, 2022 16:37
Welcome!
Posted in: Cluster BM Annotation Tips4 bp overlap in Streptomyces phages?
| posted 15 Jun, 2022 23:27
Hi Kathy,
We have a bit of evidence in mass spec data that says that the -1 start is the only one that is present. So in general, we are selecting the -1 choice over the -4 when both are present.
debbie
Posted in: Cluster BM Annotation Tips4 bp overlap in Streptomyces phages?