Assignment: Your assignment is to use your notes from class - along with help from classmates, UTAs, and me - to turn this script into a fleshed-out description of what is going on.
This is a substantial project - we’ll work on it in steps over the rest of the unit.
We are currently focused on the overall process and will cover the details over the rest of this unit.
Your first assignment is to get this script to run from top to bottom by adding all of the missing R commands. Once you have done that, you can knit it into an HTML file and upload it to RPubs. (Note - you’ll need to add the YAML header!)
Your second assignment, which will be posted later, is to answer all the TODO and other prompts to add information. You can start on this, but you don’t have to do this on your first time through the code.
Delete all the prompts like TODO() as you compete them. Use RStudio’s search function to see if you’ve missed any - there are a LOT!
Add YAML header!!! Give it a title — Title: MSA Walkthrough Assignment Output: html_document — # DONT TOUCH THE KNITR HEADER PLEASE
By: Jared H Harris
Describe how phylogeneies can be used in biology (readings will be assigned)
Make a list of at least 10 vocab terms that are important (don’t have to define) 1. Accession Number 2. Fasta Files 3. Multiple Sequence Alignment 4. Reproducible Work Flow 5. NCIB 6. Pairwise Alignment 7. Genome
8. Amino acid (single letter shortcut) 9. Mutation (types) 10. Phylogeny
Make a list of at least 5 key functions Put in the format of package::function 1. Bioconductor :: Biostrings 2. Bioconductor :: msa 3. CRAN :: rentrez 4. CRAN :: entrez_fetch 5. ggmsa :: ggmsa
Add the necessary calls to library() to load call packages Indicate which packages cam from Bioconductor, CRAN, and GitHub
# github packages
# devtools::install_github("brouwern/compbio4all")
# devtools::install_github("YuLab-SMU/ggmsa")
library(compbio4all)
library(ggmsa)
# CRAN packages
library(rentrez)
library(seqinr)
library(ape)
# Bioconductor packages
library(BiocManager)
library(Biostrings)
library(msa)
# Human shroom 3 (H. sapiens)
hShroom3 <- rentrez::entrez_fetch(db = "protein",
id = "NP_065910",
rettype = "fasta")
cat(hShroom3)
## >NP_065910.3 protein Shroom3 [Homo sapiens]
## MMRTTEDFHKPSATLNSNTATKGRYIYLEAFLEGGAPWGFTLKGGLEHGEPLIISKVEEGGKADTLSSKL
## QAGDEVVHINEVTLSSSRKEAVSLVKGSYKTLRLVVRRDVCTDPGHADTGASNFVSPEHLTSGPQHRKAA
## WSGGVKLRLKHRRSEPAGRPHSWHTTKSGEKQPDASMMQISQGMIGPPWHQSYHSSSSTSDLSNYDHAYL
## RRSPDQCSSQGSMESLEPSGAYPPCHLSPAKSTGSIDQLSHFHNKRDSAYSSFSTSSSILEYPHPGISGR
## ERSGSMDNTSARGGLLEGMRQADIRYVKTVYDTRRGVSAEYEVNSSALLLQGREARASANGQGYDKWSNI
## PRGKGVPPPSWSQQCPSSLETATDNLPPKVGAPLPPARSDSYAAFRHRERPSSWSSLDQKRLCRPQANSL
## GSLKSPFIEEQLHTVLEKSPENSPPVKPKHNYTQKAQPGQPLLPTSIYPVPSLEPHFAQVPQPSVSSNGM
## LYPALAKESGYIAPQGACNKMATIDENGNQNGSGRPGFAFCQPLEHDLLSPVEKKPEATAKYVPSKVHFC
## SVPENEEDASLKRHLTPPQGNSPHSNERKSTHSNKPSSHPHSLKCPQAQAWQAGEDKRSSRLSEPWEGDF
## QEDHNANLWRRLEREGLGQSLSGNFGKTKSAFSSLQNIPESLRRHSSLELGRGTQEGYPGGRPTCAVNTK
## AEDPGRKAAPDLGSHLDRQVSYPRPEGRTGASASFNSTDPSPEEPPAPSHPHTSSLGRRGPGPGSASALQ
## GFQYGKPHCSVLEKVSKFEQREQGSQRPSVGGSGFGHNYRPHRTVSTSSTSGNDFEETKAHIRFSESAEP
## LGNGEQHFKNGELKLEEASRQPCGQQLSGGASDSGRGPQRPDARLLRSQSTFQLSSEPEREPEWRDRPGS
## PESPLLDAPFSRAYRNSIKDAQSRVLGATSFRRRDLELGAPVASRSWRPRPSSAHVGLRSPEASASASPH
## TPRERHSVTPAEGDLARPVPPAARRGARRRLTPEQKKRSYSEPEKMNEVGIVEEAEPAPLGPQRNGMRFP
## ESSVADRRRLFERDGKACSTLSLSGPELKQFQQSALADYIQRKTGKRPTSAAGCSLQEPGPLRERAQSAY
## LQPGPAALEGSGLASASSLSSLREPSLQPRREATLLPATVAETQQAPRDRSSSFAGGRRLGERRRGDLLS
## GANGGTRGTQRGDETPREPSSWGARAGKSMSAEDLLERSDVLAGPVHVRSRSSPATADKRQDVLLGQDSG
## FGLVKDPCYLAGPGSRSLSCSERGQEEMLPLFHHLTPRWGGSGCKAIGDSSVPSECPGTLDHQRQASRTP
## CPRPPLAGTQGLVTDTRAAPLTPIGTPLPSAIPSGYCSQDGQTGRQPLPPYTPAMMHRSNGHTLTQPPGP
## RGCEGDGPEHGVEEGTRKRVSLPQWPPPSRAKWAHAAREDSLPEESSAPDFANLKHYQKQQSLPSLCSTS
## DPDTPLGAPSTPGRISLRISESVLRDSPPPHEDYEDEVFVRDPHPKATSSPTFEPLPPPPPPPPSQETPV
## YSMDDFPPPPPHTVCEAQLDSEDPEGPRPSFNKLSKVTIARERHMPGAAHVVGSQTLASRLQTSIKGSEA
## ESTPPSFMSVHAQLAGSLGGQPAPIQTQSLSHDPVSGTQGLEKKVSPDPQKSSEDIRTEALAKEIVHQDK
## SLADILDPDSRLKTTMDLMEGLFPRDVNLLKENSVKRKAIQRTVSSSGCEGKRNEDKEAVSMLVNCPAYY
## SVSAPKAELLNKIKEMPAEVNEEEEQADVNEKKAELIGSLTHKLETLQEAKGSLLTDIKLNNALGEEVEA
## LISELCKPNEFDKYRMFIGDLDKVVNLLLSLSGRLARVENVLSGLGEDASNEERSSLYEKRKILAGQHED
## ARELKENLDRRERVVLGILANYLSEEQLQDYQHFVKMKSTLLIEQRKLDDKIKLGQEQVKCLLESLPSDF
## IPKAGALALPPNLTSEPIPAGGCTFSGIFPTLTSPL
# Mouse shroom 3a (M. musculus)
mShroom3a <- entrez_fetch(db = "protein",
id = "AAF13269",
rettype = "fasta")
# Human shroom 2 (H. sapiens)
hShroom2 <- entrez_fetch(db = "protein",
id = "CAA58534",
rettype = "fasta")
# Sea-urchin shroom
sShroom <- entrez_fetch(db = "protein",
id = "XP_783573",
rettype = "fasta")
nchar(hShroom3)
## [1] 2070
nchar(mShroom3a)
## [1] 2083
nchar(sShroom)
## [1] 1758
nchar(hShroom2)
## [1] 1673
library(compbio4all)
fasta_cleaner
## function (fasta_object, parse = TRUE)
## {
## fasta_object <- sub("^(>)(.*?)(\\n)(.*)(\\n\\n)", "\\4",
## fasta_object)
## fasta_object <- gsub("\n", "", fasta_object)
## if (parse == TRUE) {
## fasta_object <- stringr::str_split(fasta_object, pattern = "",
## simplify = FALSE)
## }
## return(fasta_object[[1]])
## }
## <bytecode: 0x7ff776269608>
## <environment: namespace:compbio4all>
TODO: explain how to add the function to your R session even if you can’t download compbio4all # To add fasta_cleaner to R if you don’t have compbio4all downloaded, you can input devtools :: install_github(“brouwern/compbio4all”) which will install the file to the current R session.
fasta_cleaner <- function(fasta_object, parse = TRUE){
fasta_object <- sub("^(>)(.*?)(\\n)(.*)(\\n\\n)","\\4",fasta_object)
fasta_object <- gsub("\n", "", fasta_object)
if(parse == TRUE){
fasta_object <- stringr::str_split(fasta_object,
pattern = "",
simplify = FALSE)
}
return(fasta_object[[1]])
}
TODO: briefly explain what this code chunk is doing # This chunk of code is “cleaning up the shroom files so that the data will be properly formatted to make sequences and phylogentic trees. It is”scrubbing" our data.
hShroom3 <- fasta_cleaner(hShroom3, parse = F)
mShroom3a <- fasta_cleaner(mShroom3a, parse = F)
hShroom2 <- fasta_cleaner(hShroom2, parse = F)
sShroom <- fasta_cleaner(sShroom, parse = F)
hShroom3
## [1] "MMRTTEDFHKPSATLNSNTATKGRYIYLEAFLEGGAPWGFTLKGGLEHGEPLIISKVEEGGKADTLSSKLQAGDEVVHINEVTLSSSRKEAVSLVKGSYKTLRLVVRRDVCTDPGHADTGASNFVSPEHLTSGPQHRKAAWSGGVKLRLKHRRSEPAGRPHSWHTTKSGEKQPDASMMQISQGMIGPPWHQSYHSSSSTSDLSNYDHAYLRRSPDQCSSQGSMESLEPSGAYPPCHLSPAKSTGSIDQLSHFHNKRDSAYSSFSTSSSILEYPHPGISGRERSGSMDNTSARGGLLEGMRQADIRYVKTVYDTRRGVSAEYEVNSSALLLQGREARASANGQGYDKWSNIPRGKGVPPPSWSQQCPSSLETATDNLPPKVGAPLPPARSDSYAAFRHRERPSSWSSLDQKRLCRPQANSLGSLKSPFIEEQLHTVLEKSPENSPPVKPKHNYTQKAQPGQPLLPTSIYPVPSLEPHFAQVPQPSVSSNGMLYPALAKESGYIAPQGACNKMATIDENGNQNGSGRPGFAFCQPLEHDLLSPVEKKPEATAKYVPSKVHFCSVPENEEDASLKRHLTPPQGNSPHSNERKSTHSNKPSSHPHSLKCPQAQAWQAGEDKRSSRLSEPWEGDFQEDHNANLWRRLEREGLGQSLSGNFGKTKSAFSSLQNIPESLRRHSSLELGRGTQEGYPGGRPTCAVNTKAEDPGRKAAPDLGSHLDRQVSYPRPEGRTGASASFNSTDPSPEEPPAPSHPHTSSLGRRGPGPGSASALQGFQYGKPHCSVLEKVSKFEQREQGSQRPSVGGSGFGHNYRPHRTVSTSSTSGNDFEETKAHIRFSESAEPLGNGEQHFKNGELKLEEASRQPCGQQLSGGASDSGRGPQRPDARLLRSQSTFQLSSEPEREPEWRDRPGSPESPLLDAPFSRAYRNSIKDAQSRVLGATSFRRRDLELGAPVASRSWRPRPSSAHVGLRSPEASASASPHTPRERHSVTPAEGDLARPVPPAARRGARRRLTPEQKKRSYSEPEKMNEVGIVEEAEPAPLGPQRNGMRFPESSVADRRRLFERDGKACSTLSLSGPELKQFQQSALADYIQRKTGKRPTSAAGCSLQEPGPLRERAQSAYLQPGPAALEGSGLASASSLSSLREPSLQPRREATLLPATVAETQQAPRDRSSSFAGGRRLGERRRGDLLSGANGGTRGTQRGDETPREPSSWGARAGKSMSAEDLLERSDVLAGPVHVRSRSSPATADKRQDVLLGQDSGFGLVKDPCYLAGPGSRSLSCSERGQEEMLPLFHHLTPRWGGSGCKAIGDSSVPSECPGTLDHQRQASRTPCPRPPLAGTQGLVTDTRAAPLTPIGTPLPSAIPSGYCSQDGQTGRQPLPPYTPAMMHRSNGHTLTQPPGPRGCEGDGPEHGVEEGTRKRVSLPQWPPPSRAKWAHAAREDSLPEESSAPDFANLKHYQKQQSLPSLCSTSDPDTPLGAPSTPGRISLRISESVLRDSPPPHEDYEDEVFVRDPHPKATSSPTFEPLPPPPPPPPSQETPVYSMDDFPPPPPHTVCEAQLDSEDPEGPRPSFNKLSKVTIARERHMPGAAHVVGSQTLASRLQTSIKGSEAESTPPSFMSVHAQLAGSLGGQPAPIQTQSLSHDPVSGTQGLEKKVSPDPQKSSEDIRTEALAKEIVHQDKSLADILDPDSRLKTTMDLMEGLFPRDVNLLKENSVKRKAIQRTVSSSGCEGKRNEDKEAVSMLVNCPAYYSVSAPKAELLNKIKEMPAEVNEEEEQADVNEKKAELIGSLTHKLETLQEAKGSLLTDIKLNNALGEEVEALISELCKPNEFDKYRMFIGDLDKVVNLLLSLSGRLARVENVLSGLGEDASNEERSSLYEKRKILAGQHEDARELKENLDRRERVVLGILANYLSEEQLQDYQHFVKMKSTLLIEQRKLDDKIKLGQEQVKCLLESLPSDFIPKAGALALPPNLTSEPIPAGGCTFSGIFPTLTSPL"
align.h3.vs.m3a <- Biostrings::pairwiseAlignment(
hShroom3,
mShroom3a)
align.h3.vs.m3a
## Global PairwiseAlignmentsSingleSubject (1 of 1)
## pattern: MMRTTEDFHKPSATLN-SNTATKGRYIYLEAFLE...KAGALALPPNLTSEPIPAGGCTFSGIFPTLTSPL
## subject: MK-TPENLEEPSATPNPSRTPTE-RFVYLEALLE...KAGAISLPPALTGHATPGGTSVFGGVFPTLTSPL
## score: 2189.934
# add necessary function *No idea if this is what he wanted to be added here-check at office hours
Biostrings:: pid(align.h3.vs.m3a)
## [1] 70.56511
align.h3.vs.h2 <- Biostrings::pairwiseAlignment(
hShroom3,
hShroom2)
score(align.h3.vs.h2)
## [1] -5673.853
Biostrings::pid(align.h3.vs.h2)
## [1] 33.83277
shroom_table <- c("CAA78718" , "X. laevis Apx" , "xShroom1",
"NP_597713" , "H. sapiens APXL2" , "hShroom1",
"CAA58534" , "H. sapiens APXL", "hShroom2",
"ABD19518" , "M. musculus Apxl" , "mShroom2",
"AAF13269" , "M. musculus ShroomL" , "mShroom3a",
"AAF13270" , "M. musculus ShroomS" , "mShroom3b",
"NP_065910", "H. sapiens Shroom" , "hShroom3",
"ABD59319" , "X. laevis Shroom-like", "xShroom3",
"NP_065768", "H. sapiens KIAA1202" , "hShroom4a",
"AAK95579" , "H. sapiens SHAP-A" , "hShroom4b",
#"DQ435686" , "M. musculus KIAA1202" , "mShroom4",
"ABA81834" , "D. melanogaster Shroom", "dmShroom",
"EAA12598" , "A. gambiae Shroom", "agShroom",
"XP_392427" , "A. mellifera Shroom" , "amShroom",
"XP_783573" , "S. purpuratus Shroom" , "spShroom") #sea urchin
# convert to Matrix
shroom_table_matrix <- matrix(shroom_table,
byrow = T,
nrow = 14)
# convert to Data Frame
shroom_table <- data.frame(shroom_table_matrix,
stringsAsFactors = F)
# Naming Columns
names(shroom_table) <- c("accession", "name.orig","name.new")
# Create simplified species names
shroom_table$spp <- "Homo"
shroom_table$spp[grep("laevis",shroom_table$name.orig)] <- "Xenopus"
shroom_table$spp[grep("musculus",shroom_table$name.orig)] <- "Mus"
shroom_table$spp[grep("melanogaster",shroom_table$name.orig)] <- "Drosophila"
shroom_table$spp[grep("gambiae",shroom_table$name.orig)] <- "mosquito"
shroom_table$spp[grep("mellifera",shroom_table$name.orig)] <- "bee"
shroom_table$spp[grep("purpuratus",shroom_table$name.orig)] <- "sea urchin"
shroom_table
## accession name.orig name.new spp
## 1 CAA78718 X. laevis Apx xShroom1 Xenopus
## 2 NP_597713 H. sapiens APXL2 hShroom1 Homo
## 3 CAA58534 H. sapiens APXL hShroom2 Homo
## 4 ABD19518 M. musculus Apxl mShroom2 Mus
## 5 AAF13269 M. musculus ShroomL mShroom3a Mus
## 6 AAF13270 M. musculus ShroomS mShroom3b Mus
## 7 NP_065910 H. sapiens Shroom hShroom3 Homo
## 8 ABD59319 X. laevis Shroom-like xShroom3 Xenopus
## 9 NP_065768 H. sapiens KIAA1202 hShroom4a Homo
## 10 AAK95579 H. sapiens SHAP-A hShroom4b Homo
## 11 ABA81834 D. melanogaster Shroom dmShroom Drosophila
## 12 EAA12598 A. gambiae Shroom agShroom mosquito
## 13 XP_392427 A. mellifera Shroom amShroom bee
## 14 XP_783573 S. purpuratus Shroom spShroom sea urchin
$ allows us to call up a specific column within the table and display it in R by itself. In this case, the output is the “accession” column.shroom_table$accession
## [1] "CAA78718" "NP_597713" "CAA58534" "ABD19518" "AAF13269" "AAF13270"
## [7] "NP_065910" "ABD59319" "NP_065768" "AAK95579" "ABA81834" "EAA12598"
## [13] "XP_392427" "XP_783573"
# add necessary function *I think this is right
shrooms <- rentrez :: entrez_fetch(db = "protein",
id = shroom_table$accession,
rettype = "fasta")
cat(shrooms)
TODO: in a brief sentence explain what this is doing and if/how its different from the previous code chunks # This code chunk is using the entrez_fetch_list function to_____ and it is different from the previous chunks because it —
shrooms_list <- entrez_fetch_list(db = "protein",
id = shroom_table$accession,
rettype = "fasta")
length(shrooms_list)
## [1] 14
for(i in 1:length(shrooms_list)){
shrooms_list[[i]] <- fasta_cleaner(shrooms_list[[i]], parse = F)
}
TODO: summarize what is going on in this code chunk, then annotate each line of code with what its doing # The rep() function is replicating the _______. The for() function is used to repeat a task mulitple times, and in this instance it is ______. The names() function is being used to add names to _____/
# Replication
shrooms_vector <- rep(NA, length(shrooms_list))
# For loop cleaning
for(i in 1:length(shrooms_vector)){
shrooms_vector[i] <- shrooms_list[[i]]
}
# Adding names
names(shrooms_vector) <- names(shrooms_list)
#
shrooms_vector_ss <- Biostrings:: AAStringSet (shrooms_vector)
shrooms_align <- msa(shrooms_vector_ss,
method = "ClustalW")
## use default substitution matrix
shrooms_align
## CLUSTAL 2.1
##
## Call:
## msa(shrooms_vector_ss, method = "ClustalW")
##
## MsaAAMultipleAlignment with 14 rows and 2252 columns
## aln names
## [1] -------------------------...------------------------- NP_065768
## [2] -------------------------...------------------------- AAK95579
## [3] -------------------------...SVFGGVFPTLTSPL----------- AAF13269
## [4] -------------------------...SVFGGVFPTLTSPL----------- AAF13270
## [5] -------------------------...CTFSGIFPTLTSPL----------- NP_065910
## [6] -------------------------...NKS--LPPPLTSSL----------- ABD59319
## [7] -------------------------...------------------------- CAA58534
## [8] -------------------------...------------------------- ABD19518
## [9] -------------------------...LT----------------------- NP_597713
## [10] -------------------------...------------------------- CAA78718
## [11] -------------------------...------------------------- EAA12598
## [12] -------------------------...------------------------- ABA81834
## [13] MTELQPSPPGYRVQDEAPGPPSCPP...------------------------- XP_392427
## [14] -------------------------...AATSSSSNGIGGPEQLNSNATSSYC XP_783573
## Con -------------------------...------------------------- Consensus
TODO: briefly explain what is being done in this chunk. This is tricky (and annoying) so do your best
# WHAT IS THE LINE BELOW DOING? (its tricky - do your best)
class(shrooms_align) <- "AAMultipleAlignment"
# WHAT IS THE LINE BELOW DOING? This is simpler
shrooms_align_seqinr <- msaConvert(shrooms_align, type = "seqinr::alignment")
TODO: what is the output this produces # The output of this is an ugly version of the MSA.
print_msa(alignment = shrooms_align_seqinr,
chunksize = 60)
library(ggmsa)
ggmsa :: ggmsa(shrooms_align, # shrooms_align, NOT shrooms_align_seqinr
start = 2000,
end = 2100)
msa::msaPrettyPrint(shrooms_align, # alignment
file = "shroom_msa.pdf", # file name
y=c(2000, 2100), # range
askForOverwrite=FALSE)
getwd()
## [1] "/Users/jaredharris/Downloads"