```{bash} cd /home/shared/8TB_HDD_02/cnmntgna/Applications/bioinfo/ curl -O https://ftp.ncbi.nlm.nih.gov/blast/executables/blast+/LATEST/ncbi-blast-2.13.0+-x64-linux.tar.gz tar -xf ncbi-blast-2.13.0+-x64-linux.tar.gz ``` ```{bash} ls /home/shared/8TB_HDD_02/cnmntgna/Applications/bioinfo/ # trying to remove macos copy, receive an error message # made the removal happen over in the 'Terminal' access tab # rm -r ncbi-blast-2.13.0+-x64-macosx.tar.gz ``` ```{bash} /home/shared/8TB_HDD_02/cnmntgna/Applications/bioinfo/ncbi-blast-2.13.0+/bin/blastx -h ``` ```{bash} cd ../data curl -O https://ftp.uniprot.org/pub/databases/uniprot/current_release/knowledgebase/complete/uniprot_sprot.fasta.gz mv uniprot_sprot.fasta.gz uniprot_sprot_r2023_01.fasta.gz gunzip -k uniprot_sprot_r2023_01.fasta.gz ls ../data ``` ```{bash} # add r engine/ false echo code for rendering in 1.1 /home/shared/8TB_HDD_02/cnmntgna/Applications/bioinfo/ncbi-blast-2.13.0+/bin/makeblastdb \ -in /home/shared/8TB_HDD_02/cnmntgna/data/uniprot_sprot_r2023_01.fasta \ -dbtype prot \ -out /home/shared/8TB_HDD_02/cnmntgna/output/blastdb/uniprot_sprot_r2023_01 ``` ```{bash} curl https://eagle.fish.washington.edu/cnidarian/Ab_4denovo_CLC6_a.fa \ -k \ > /home/shared/8TB_HDD_02/cnmntgna/data/Ab_4denovo_CLC6_a.fa ``` ```{bash} head /home/shared/8TB_HDD_02/cnmntgna/data/Ab_4denovo_CLC6_a.fa echo "How many sequences are there?" grep -c ">" /home/shared/8TB_HDD_02/cnmntgna/data/Ab_4denovo_CLC6_a.fa ``` ```{bash} #fix my paths for reproducibility purposes, the full path is incorrect for that purpose /home/shared/8TB_HDD_02/cnmntgna/Applications/bioinfo/ncbi-blast-2.13.0+/bin/blastx \ -query /home/shared/8TB_HDD_02/cnmntgna/data/Ab_4denovo_CLC6_a.fa \ -db /home/shared/8TB_HDD_02/cnmntgna/output/blastdb/uniprot_sprot_r2023_01 \ -out /home/shared/8TB_HDD_02/cnmntgna/output/Ab_4-uniprot_blastx.tab \ -evalue 1E-20 \ -num_threads 20 \ -max_target_seqs 1 \ -outfmt 6 #code throws the following warning: Warning: [blastx] Examining 5 or more matches is recommended ``` ```{bash} head -2 /home/shared/8TB_HDD_02/cnmntgna/output/Ab_4-uniprot_blastx.tab wc -l /home/shared/8TB_HDD_02/cnmntgna/output/Ab_4-uniprot_blastx.tab ``` ```{bash} curl -O "Accept: text/plain; format=tsv" "https://rest.uniprot.org/uniprotkb/search?query=reviewed:true+AND+organism_id:9606" ``` ```{bash} #THIS CODE CAUSES ERROR curl -O -H "Accept: text/plain; format=tsv" "https://rest.uniprot.org/uniprotkb/stream?compressed=true&fields=accession%2Creviewed%2Cid%2Cprotein_name%2Cgene_names%2Corganism_name%2Clength%2Cgo_f%2Cgo%2Cgo_p%2Cgo_c%2Cgo_id%2Ccc_interaction%2Cec%2Cxref_reactome%2Cxref_unipathway%2Cxref_interpro&format=tsv&query=%28%2A%29%20AND%20%28reviewed%3Atrue%29" #this is the reference table with GO terms and protein names to join with my blast output terms. I can use the second gannet url to bypass. This is an easy confuse becasue my first DB is named the same but is just so I can reference. Note I am aligning, not comparing the sequences. ``` ```{bash} head -2 /home/shared/8TB_HDD_02/cnmntgna/output/Ab_4-uniprot_blastx.tab wc -l /home/shared/8TB_HDD_02/cnmntgna/output/Ab_4-uniprot_blastx.tab ``` ```{bash} #not sure what broken pipe means, google says it is a mismatch of tasks, but i need to look further to understand what that means. tr '|' '\t' < /home/shared/8TB_HDD_02/cnmntgna/output/Ab_4-uniprot_blastx.tab | head -2 ``` ```{bash} tr '|' '\t' < /home/shared/8TB_HDD_02/cnmntgna/output/Ab_4-uniprot_blastx.tab \ > /home/shared/8TB_HDD_02/cnmntgna/output/Ab_4-uniprot_blastx_sep.tab ``` ```{bash} head -2 /home/shared/8TB_HDD_02/cnmntgna/data/uniprot_table_r2023_01.tab wc -l /home/shared/8TB_HDD_02/cnmntgna/data/uniprot_table_r2023_01.tab ``` ```{r} library(tidyverse) install.packages("kableExtra") library("kableExtra") ``` ```{r} bltabl <- read.csv("../output/Ab_4-uniprot_blastx_sep.tab", sep = '\t', header = FALSE) spgo <- read.csv("../data/uniprot_table_r2023_01.tab", sep = '\t', header = TRUE) str(spgo) ``` ```{r} kbl( head( left_join(bltabl, spgo, by = c("V3" = "Entry")) %>% select(V1, V3, V13, Protein.names, Organism, Gene.Ontology..biological.process., Gene.Ontology.IDs) %>% mutate(V1 = str_replace_all(V1, pattern = "solid0078_20110412_FRAG_BC_WHITE_WHITE_F3_QV_SE_trimmed", replacement = "Ab")) ) ) %>% kable_styling(bootstrap_options = c("striped", "hover", "condensed", "responsive")) ``` ```{r} left_join(bltabl, spgo, by = c("V3" = "Entry")) %>% select(V1, V3, V13, Protein.names, Organism, Gene.Ontology..biological.process., Gene.Ontology.IDs) %>% mutate(V1 = str_replace_all(V1, pattern = "solid0078_20110412_FRAG_BC_WHITE_WHITE_F3_QV_SE_trimmed", replacement = "Ab")) %>% write_delim("../output/blast_annot_go.tab", delim = '\t') ``` ```{r} annot_tab <- read.csv("../output/blast_annot_go.tab", sep = '\t', header = TRUE) ```