Mitochondrial Genome Annotation
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1 Protein Genes 1,2 1 Institute of Bioinformatics University of Leipzig 2 Department of Bioinformatics Lebanese University TBI Bled 2015
2 Outline Introduction Mitochondrial DNA Problem Tools Training Annotation
3 Mitochondrial DNA Problem Mitochondrial DNA Circular molecule located in mitochondria within eurkaryotic cells. transform energy to a form used by the cells Length about nucleotide. 13 protein coding genes 22 trna genes 2 rrna [wikipedia]
4 Mitochondrial DNA Problem Problem Refseq is the most used repository for mitochondrial genome annotation Refseq suffers form several inconsistencies and errors in annotation Missing or incorrect strand Confusing trnl1/trnl2 trns1/trns2 Inconsistencies in gene names(bernt et. al 2012) Problem in developing automated analysis for mitochondrial data
5 Mitochondrial DNA Problem Objective Develop an automated pipeline for mtdna annotation by refining taxon specific hmm and covariance models.
6 Tools Training Annotation Tools HMMER an implementation of profile HMMs (Sean Eddy and his group ). hmmbuild build a model from multiple sequences hmmalign algin a model to sequences hmmsearch search a model in sequences database hmmscan search a genome in models database
7 Tools Training Annotation Training Data Build taxon specific models along the phylogenetic tree nodes Step 1: Build protein models for leaf sequences ABCD AB A B C Step 2: Recursively lift up the model with best score CD D Step 3: Build the models database
8 Tools Training Annotation Training Data Build taxon specific models along the phylogenetic tree nodes Step 1: Build protein models for leaf sequences ABCD AB M A A M B B M C C Step 2: Recursively lift up the model with best score CD M D D Step 3: Build the models database
9 Tools Training Annotation Training Data Build taxon specific models along the phylogenetic tree nodes Step 1: Build protein models for leaf sequences Step 2: Recursively lift up the model with best score ABCD M AB M CD AB CD M A A M B B M C C M D D Step 3: Build the models database
10 Tools Training Annotation Training Data Build taxon specific models along the phylogenetic tree nodes Step 1: Build protein models for leaf sequences Step 2: Recursively lift up the model with best score Step 3: Build the models database M AB M ABCD ABCD M CD AB CD M A A M B B M C C M D D
11 Tools Training Annotation Example: Part of nad1 Alignment
12 Tools Training Annotation Annotation Input Start Fasta Translate in 6 Frames Taxonomic level database Search DB Overlap no Output Bed yes Best hit
13 Train 3843 mt genome sequence from refseq63 Test on 925 genome which are newly annotated in refseq69 Scan against level models database
14 Phylum and Class Models Phylum models
15 Phylum and Class Models Phylum models Class models
16 Phylum and Class Models Sn = TP TP+FN Sp = TP TP+FP Phylum models Class models
17 Phylum and Class Models Sn = TP TP+FN Sp = TP TP+FP Phylum models Class models Sn = Sp = Sn = Sp = 0.988
18 An automated pipeline to annotate protein coding genes in mtdna by refining taxon specific hmm Outlook Find the best level database and best parameters to minimize FN and maximize TP Analyse the results in deep to improve the refseq annotation Apply on trna
19 Thanks to Matthias Bernt Christian Honer Frank Juhling Abdullah Sahyoun Peter Stadler
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