The goal of this exercise is to make you familiar with how to download data from Google Sheets and to briefly review some key concepts R functions and coding concepts.
We’ll do the following things
(TODO: MAKE YOUR OWN OUTLINE)
## Google sheets download package
# comment this out when you are done
#install.packages("googlesheets4")
library(googlesheets4)
# comp bio packages
library(seqinr)
library(rentrez)
library(compbio4all)
library(Biostrings)
## Loading required package: BiocGenerics
##
## Attaching package: 'BiocGenerics'
## The following objects are masked from 'package:stats':
##
## IQR, mad, sd, var, xtabs
## The following objects are masked from 'package:base':
##
## anyDuplicated, append, as.data.frame, basename, cbind, colnames,
## dirname, do.call, duplicated, eval, evalq, Filter, Find, get, grep,
## grepl, intersect, is.unsorted, lapply, Map, mapply, match, mget,
## order, paste, pmax, pmax.int, pmin, pmin.int, Position, rank,
## rbind, Reduce, rownames, sapply, setdiff, sort, table, tapply,
## union, unique, unsplit, which.max, which.min
## Loading required package: S4Vectors
## Warning: package 'S4Vectors' was built under R version 4.1.2
## Loading required package: stats4
##
## Attaching package: 'S4Vectors'
## The following objects are masked from 'package:base':
##
## expand.grid, I, unname
## Loading required package: IRanges
## Loading required package: XVector
## Loading required package: GenomeInfoDb
##
## Attaching package: 'Biostrings'
## The following object is masked from 'package:seqinr':
##
## translate
## The following object is masked from 'package:base':
##
## strsplit
WHAT IS THIS? THIS IS THE SPREADSHEET OF OUR GENES, AND WE ARE USING A WEB ADDRESS FOR OUR DATA
spreadsheet_sp <- "https://docs.google.com/spreadsheets/d/1spC_ZA3_cVuvU3e_Jfcj2nEIfzp-vaP7SA5f-qwQ1pg/edit?usp=sharing"
WHAT DOES THIS DO? THIS IS MAKING SURE THERE ARE NO CREDENTIALS NECESSART
# be sure to run this!
googlesheets4::gs4_deauth() # <====== MUST RUN THIS
Third, we download our data.
“Error in curl::curl_fetch_memory(url, handle = handle) : Error in the HTTP2 framing layer”
If that happens, just re-run the code.
# I include this again in case you missed is the first time : )
googlesheets4::gs4_deauth()
# download
## NOTE: if you get an error, just run the code again
refseq_column <- read_sheet(ss = spreadsheet_sp, # the url
sheet = "RefSeq_prot", # the name of the worksheet
range = "selenoprot!H1:H364",
col_names = TRUE,
na = "", # fill in empty spaces "" w/NA
trim_ws = TRUE)
## ✓ Reading from "human_gene_table".
## ✓ Range ''selenoprot'!H1:H364'.
## NOTE: if you get an error, just run the code again
# for reasons we won't get into I'm going to do this
protein_refseq <- refseq_column$RefSeq_prot
WHAT’S THIS? THIS IS GETTING THE REFERENCE SEQUENCES OF THE FIRST 10
protein_refseq[1:10]
## [1] "NP_000783.2" "NP_998758.1" "NP_001034804.1" "NP_001034805.1"
## [5] "NP_001311245.1" NA NA "NP_054644.1"
## [9] "NP_001353425.1" "NP_000784.3"
WHAT’S THIS? THIS IS DOWNLOADING THE RANGE OF CELLS IN THE SPREADSHEET GENE
# download
## NOTE: if you get an error, just run the code again
gene_name_column <- read_sheet(ss = spreadsheet_sp, # the url
sheet = "gene", # the name of the worksheet
range = "selenoprot!A1:A364",
col_names = TRUE,
na = "", # fill in empty spaces "" w/NA
trim_ws = TRUE)
## ✓ Reading from "human_gene_table".
## ✓ Range ''selenoprot'!A1:A364'.
## NOTE: if you get an error, just run the code again
# for reasons we won't get into I'm going to do this
gene <- gene_name_column$gene
WHAT’S THIS DOING DOWNLOADING A VECTOR OF CHARACTER DATA
is(protein_refseq)
## [1] "character" "vector"
## [3] "data.frameRowLabels" "SuperClassMethod"
## [5] "character_OR_connection" "character_OR_NULL"
## [7] "atomic" "EnumerationValue"
## [9] "vector_OR_Vector" "vector_OR_factor"
class(protein_refseq)
## [1] "character"
length(protein_refseq)
## [1] 363
protein_refseq[1:10]
## [1] "NP_000783.2" "NP_998758.1" "NP_001034804.1" "NP_001034805.1"
## [5] "NP_001311245.1" NA NA "NP_054644.1"
## [9] "NP_001353425.1" "NP_000784.3"
WHAT’S THIS DOING? CHECKING FOR THE NA’S
is.na(protein_refseq)
## [1] FALSE FALSE FALSE FALSE FALSE TRUE TRUE FALSE FALSE FALSE FALSE TRUE
## [13] TRUE FALSE FALSE FALSE FALSE FALSE FALSE FALSE TRUE TRUE TRUE FALSE
## [25] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE TRUE TRUE FALSE FALSE
## [37] FALSE TRUE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [49] FALSE FALSE FALSE TRUE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [61] FALSE FALSE FALSE FALSE FALSE FALSE FALSE TRUE FALSE FALSE FALSE FALSE
## [73] TRUE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE TRUE
## [85] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [97] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [109] FALSE TRUE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [121] TRUE FALSE TRUE FALSE FALSE FALSE FALSE FALSE FALSE FALSE TRUE FALSE
## [133] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [145] FALSE FALSE FALSE FALSE TRUE FALSE FALSE FALSE FALSE FALSE FALSE TRUE
## [157] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [169] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [181] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [193] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE TRUE TRUE FALSE
## [205] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [217] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [229] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [241] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE TRUE TRUE
## [253] FALSE FALSE FALSE TRUE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [265] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [277] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE TRUE FALSE
## [289] TRUE TRUE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [301] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [313] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [325] FALSE FALSE FALSE FALSE FALSE FALSE TRUE FALSE FALSE FALSE FALSE FALSE
## [337] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [349] FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE FALSE
## [361] FALSE FALSE FALSE
WHAT’S THIS DOING? MAKING A TABLE OF THE NA’S AND NOT, FALSE IN THIS SCENARIO MEANS NA
table(is.na(protein_refseq))
##
## FALSE TRUE
## 334 29
WHAT’S THIS DOING? THIS IS JUST SHOWING ANOTHER WAY OF SEEING HOW MANY NA’S WE HAVE
# ...
temp <- is.na(protein_refseq)
# ....
protein_refseq[temp]
## [1] NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA NA
## [26] NA NA NA NA
temp2 <- protein_refseq[temp]
# ...
length(temp2)
## [1] 29
WHAT’S THIS DOING? WNY? MAKING A DATA FRAME WITH GENE AND THE PROTEIN_REFSEQ
seleno_df <- data.frame(gene = gene,
protein_refseq = protein_refseq)
WHAT’S THIS DOING? MAKING A SUMMARY
summary(seleno_df)
## gene protein_refseq
## Length:363 Length:363
## Class :character Class :character
## Mode :character Mode :character
head(seleno_df)
## gene protein_refseq
## 1 DIO1 NP_000783.2
## 2 DIO1 NP_998758.1
## 3 DIO1 NP_001034804.1
## 4 DIO1 NP_001034805.1
## 5 DIO1 NP_001311245.1
## 6 DIO1 <NA>
WHAT’S THIS DOING? CAN TAKE OUT ALL THE NA’S USING NA.OMIT
# omit NAs
seleno_df_noNA <- na.omit(seleno_df)
# check length- should be shorter
dim(seleno_df)
## [1] 363 2
dim(seleno_df_noNA)
## [1] 334 2
The same gene can appear multiple times because multiple isoforms are listed.
head(seleno_df_noNA)
## gene protein_refseq
## 1 DIO1 NP_000783.2
## 2 DIO1 NP_998758.1
## 3 DIO1 NP_001034804.1
## 4 DIO1 NP_001034805.1
## 5 DIO1 NP_001311245.1
## 8 DIO2 NP_054644.1
WHAT’S THIS DOING? USING THE FUNCTION UNIQUE() TO SELECT ONE ROW FOR EACH GENE
genes_unique <- unique(seleno_df_noNA$gene)
length(genes_unique)
## [1] 37
genes_unique
## [1] "DIO1" "DIO2" "DIO3" "GPX1" "GPX2" "GPX3"
## [7] "GPX4" "GPX6" "MSRB1" "SELENOF" "SELENOH" "SELENOI"
## [13] "SELENOK" "SELENOM" "SELENON" "SELENOO" "SELENOP" "SELENOS"
## [19] "SELENOT" "SELENOV" "SELENOW" "SEPHS2" "TXNRD1" "TXNRD2"
## [25] "TXNRD3" "SELENOP1" "SELENOP2" "SELENOU" "SELENOW1" "SELENOW2"
## [31] "SELENOE" "SELENOJ" "SELENOL" "SELENOO1" "SELENOO2" "SELENOT1"
## [37] "SELENOT2"
unique() just gives us the unique elements. A related function, duplicated(), gives us the location of duplicated elements in the vector. FALSE means “not duplicated yet” or “first instance so far”.
i.dups <- duplicated(seleno_df_noNA$gene)
We can remove the duplicates using a form of reverse indexing where the “!” means “not”. (You don’t need to know this for the exam)
seleno_df_noNA[!i.dups, ]
## gene protein_refseq
## 1 DIO1 NP_000783.2
## 8 DIO2 NP_054644.1
## 14 DIO3 NP_001353.4
## 15 GPX1 NP_000572.2
## 20 GPX2 NP_002074.2
## 24 GPX3 NP_002075.2
## 26 GPX4 NP_002076.2
## 29 GPX6 NP_874360.1
## 30 MSRB1 NP_057416.1
## 31 SELENOF NP_004252.2
## 35 SELENOH NP_734467.1
## 37 SELENOI NP_277040.1
## 39 SELENOK NP_067060.2
## 40 SELENOM NP_536355.1
## 41 SELENON NP_996809.1
## 43 SELENOO NP_113642.1
## 44 SELENOP NP_005401.3
## 47 SELENOS NP_060915.2
## 49 SELENOT NP_057359.2
## 50 SELENOV NP_874363.1
## 53 SELENOW NP_003000.1
## 54 SEPHS2 NP_036380.2
## 55 TXNRD1 NP_877393.1
## 62 TXNRD2 NP_006431.2
## 69 TXNRD3 NP_443115.1
## 232 SELENOP1 NP_001026780.2
## 233 SELENOP2 NP_001335698.1
## 236 SELENOU NP_001180447.1
## 268 SELENOW1 NP_001291715.2
## 269 SELENOW2 NP_001341647.1
## 334 SELENOE NP_001182713.2
## 338 SELENOJ NP_001180398.1
## 340 SELENOL NP_001177311.1
## 343 SELENOO1 NP_001038336.2
## 344 SELENOO2 NP_001335014.1
## 348 SELENOT1 NP_840075.2
## 350 SELENOT2 NP_001091957.2
Make a dataframe of non-duplicated genes
seleno_df_noDups <- seleno_df_noNA[!i.dups, ]
dim(seleno_df_noDups)
## [1] 37 2
Let’s select 2 random sequences to work with. We’ll use WHICH FUNCTION? to select a random index number to get
First, lets make a vector that contains a unique number for each row of data
indices <- 1:nrow(seleno_df_noDups)
This would do the same thing
# with dim
indices <- 1:dim(seleno_df_noDups)[1]
# with length
indices <- 1:length(seleno_df_noDups$gene)
or hard-coded
indices <- 1:37
We can then use WHICH FUNCTION? to select 2 random numbers from this vector.
For x = we’ll use our vector of indices (1 to 37). For size we’ll use 2, since we want to pull out just 2 numbers. For replace we’ll use WHAT? since we don’t want to be ale to select the same number twice.
i.random.genes <- sample(x = indices,
size = 2,
replace = FALSE)
Hard coded this would be
i.random.genes <- sample(x = c(1:37),
size = 2,
replace = FALSE)
This gives me HOW MANY? indices values. 25, 27
i.random.genes
## [1] 12 20
I can now use these index values to pull out HOW MANY? 2 rows of data
seleno_df_noNA[i.random.genes, ]
## gene protein_refseq
## 16 GPX1 NP_958799.1
## 27 GPX4 NP_001034936.1
Hard coded, this would be something like this for whichever genes happen to have been selected
seleno_df_noNA[c(37,15), ]
## gene protein_refseq
## 47 SELENOS NP_060915.2
## 19 GPX1 NP_001316384.1
I will now… DO WHAT? DOWNLOAD THE TWO FASTA FILES
rentrez::entrez_fetch(id = "NP_060915.2",
db = "protein",
rettype = "fasta")
## [1] ">NP_060915.2 selenoprotein S isoform 1 [Homo sapiens]\nMERQEESLSARPALETEGLRFLHTTVGSLLATYGWYIVFSCILLYVVFQKLSARLRALRQRQLDRAAAAV\nEPDVVVKRQEALAAARLKMQEELNAQVEKHKEKLKQLEEEKRRQKIEMWDSMQEGKSYKGNAKKPQEEDS\nPGPSTSSVLKRKSDRKPLRGGGYNPLSGEGGGACSWRPGRRGPSSGGUG\n\n"
rentrez::entrez_fetch(id = "NP_001316384.1",
db = "protein",
rettype = "fasta")
## [1] ">NP_001316384.1 glutathione peroxidase 1 isoform 5 [Homo sapiens]\nMCAARLAAAAAAAQSVYAFSARPLAGGEPVSLGSLRGKENAKNEEILNSLKYVRPGGGFEPNFMLFEKCE\nVNGAGAHPLFAFLREALPAPSDDATALMTDPKLITWSPVCRNDVAWNFEKFLVGPDGVPLRRYSRRFQTI\nDIEPDIEALLSQGPSCA\n\n"
WHAT"S THIS DOING? SAVING THEM INTO VECTORS
prot1 <- rentrez::entrez_fetch(id = "NP_060915.2",
db = "protein",
rettype = "fasta")
prot2 <- rentrez::entrez_fetch(id = "NP_001316384.1",
db = "protein",
rettype = "fasta")
I can put them into a WHAT? like this SAVING THEM INTO A VECTOR
# make the WHAT?
seleno_thingy <- vector("list", 1)
# add the first fasta
seleno_thingy[[1]] <- prot1
# See the result
seleno_thingy
## [[1]]
## [1] ">NP_060915.2 selenoprotein S isoform 1 [Homo sapiens]\nMERQEESLSARPALETEGLRFLHTTVGSLLATYGWYIVFSCILLYVVFQKLSARLRALRQRQLDRAAAAV\nEPDVVVKRQEALAAARLKMQEELNAQVEKHKEKLKQLEEEKRRQKIEMWDSMQEGKSYKGNAKKPQEEDS\nPGPSTSSVLKRKSDRKPLRGGGYNPLSGEGGGACSWRPGRRGPSSGGUG\n\n"
# add the first fasta
seleno_thingy[[2]] <- prot2
# see the result
seleno_thingy
## [[1]]
## [1] ">NP_060915.2 selenoprotein S isoform 1 [Homo sapiens]\nMERQEESLSARPALETEGLRFLHTTVGSLLATYGWYIVFSCILLYVVFQKLSARLRALRQRQLDRAAAAV\nEPDVVVKRQEALAAARLKMQEELNAQVEKHKEKLKQLEEEKRRQKIEMWDSMQEGKSYKGNAKKPQEEDS\nPGPSTSSVLKRKSDRKPLRGGGYNPLSGEGGGACSWRPGRRGPSSGGUG\n\n"
##
## [[2]]
## [1] ">NP_001316384.1 glutathione peroxidase 1 isoform 5 [Homo sapiens]\nMCAARLAAAAAAAQSVYAFSARPLAGGEPVSLGSLRGKENAKNEEILNSLKYVRPGGGFEPNFMLFEKCE\nVNGAGAHPLFAFLREALPAPSDDATALMTDPKLITWSPVCRNDVAWNFEKFLVGPDGVPLRRYSRRFQTI\nDIEPDIEALLSQGPSCA\n\n"
# WHAT DOES THIS DO?
names(seleno_thingy) <- c("prot1", "prot2")
#Output
seleno_thingy
## $prot1
## [1] ">NP_060915.2 selenoprotein S isoform 1 [Homo sapiens]\nMERQEESLSARPALETEGLRFLHTTVGSLLATYGWYIVFSCILLYVVFQKLSARLRALRQRQLDRAAAAV\nEPDVVVKRQEALAAARLKMQEELNAQVEKHKEKLKQLEEEKRRQKIEMWDSMQEGKSYKGNAKKPQEEDS\nPGPSTSSVLKRKSDRKPLRGGGYNPLSGEGGGACSWRPGRRGPSSGGUG\n\n"
##
## $prot2
## [1] ">NP_001316384.1 glutathione peroxidase 1 isoform 5 [Homo sapiens]\nMCAARLAAAAAAAQSVYAFSARPLAGGEPVSLGSLRGKENAKNEEILNSLKYVRPGGGFEPNFMLFEKCE\nVNGAGAHPLFAFLREALPAPSDDATALMTDPKLITWSPVCRNDVAWNFEKFLVGPDGVPLRRYSRRFQTI\nDIEPDIEALLSQGPSCA\n\n"
Elements of the list are accessed like this
seleno_thingy[[1]]
## [1] ">NP_060915.2 selenoprotein S isoform 1 [Homo sapiens]\nMERQEESLSARPALETEGLRFLHTTVGSLLATYGWYIVFSCILLYVVFQKLSARLRALRQRQLDRAAAAV\nEPDVVVKRQEALAAARLKMQEELNAQVEKHKEKLKQLEEEKRRQKIEMWDSMQEGKSYKGNAKKPQEEDS\nPGPSTSSVLKRKSDRKPLRGGGYNPLSGEGGGACSWRPGRRGPSSGGUG\n\n"
I’ll clean them with fasta_cleaner()
# first, make a copy of the list for storing the clean data
## I'm just going to copy over the old data
seleno_thingy_clean <- seleno_thingy
# HOW TO MAKE THIS MORE COMPACT?
for(i in 1:length(seleno_thingy_clean)){
clean_fasta_temp <- compbio4all::fasta_cleaner(seleno_thingy[[i]],
parse = T)
seleno_thingy_clean[[i]] <- clean_fasta_temp
}
Now the data looks like this HOW WOULD YOU DESCRIBE THIS? THE DATA IS SPREAD OUT, EACH ELEMENT OF THE LIST IS A VECTOR OF CHARACTER DATA
seleno_thingy_clean
## $prot1
## [1] "M" "E" "R" "Q" "E" "E" "S" "L" "S" "A" "R" "P" "A" "L" "E" "T" "E" "G"
## [19] "L" "R" "F" "L" "H" "T" "T" "V" "G" "S" "L" "L" "A" "T" "Y" "G" "W" "Y"
## [37] "I" "V" "F" "S" "C" "I" "L" "L" "Y" "V" "V" "F" "Q" "K" "L" "S" "A" "R"
## [55] "L" "R" "A" "L" "R" "Q" "R" "Q" "L" "D" "R" "A" "A" "A" "A" "V" "E" "P"
## [73] "D" "V" "V" "V" "K" "R" "Q" "E" "A" "L" "A" "A" "A" "R" "L" "K" "M" "Q"
## [91] "E" "E" "L" "N" "A" "Q" "V" "E" "K" "H" "K" "E" "K" "L" "K" "Q" "L" "E"
## [109] "E" "E" "K" "R" "R" "Q" "K" "I" "E" "M" "W" "D" "S" "M" "Q" "E" "G" "K"
## [127] "S" "Y" "K" "G" "N" "A" "K" "K" "P" "Q" "E" "E" "D" "S" "P" "G" "P" "S"
## [145] "T" "S" "S" "V" "L" "K" "R" "K" "S" "D" "R" "K" "P" "L" "R" "G" "G" "G"
## [163] "Y" "N" "P" "L" "S" "G" "E" "G" "G" "G" "A" "C" "S" "W" "R" "P" "G" "R"
## [181] "R" "G" "P" "S" "S" "G" "G" "U" "G"
##
## $prot2
## [1] "M" "C" "A" "A" "R" "L" "A" "A" "A" "A" "A" "A" "A" "Q" "S" "V" "Y" "A"
## [19] "F" "S" "A" "R" "P" "L" "A" "G" "G" "E" "P" "V" "S" "L" "G" "S" "L" "R"
## [37] "G" "K" "E" "N" "A" "K" "N" "E" "E" "I" "L" "N" "S" "L" "K" "Y" "V" "R"
## [55] "P" "G" "G" "G" "F" "E" "P" "N" "F" "M" "L" "F" "E" "K" "C" "E" "V" "N"
## [73] "G" "A" "G" "A" "H" "P" "L" "F" "A" "F" "L" "R" "E" "A" "L" "P" "A" "P"
## [91] "S" "D" "D" "A" "T" "A" "L" "M" "T" "D" "P" "K" "L" "I" "T" "W" "S" "P"
## [109] "V" "C" "R" "N" "D" "V" "A" "W" "N" "F" "E" "K" "F" "L" "V" "G" "P" "D"
## [127] "G" "V" "P" "L" "R" "R" "Y" "S" "R" "R" "F" "Q" "T" "I" "D" "I" "E" "P"
## [145] "D" "I" "E" "A" "L" "L" "S" "Q" "G" "P" "S" "C" "A"
HOW WOULD YOU DESCRIBE THIS? MAKING SURE THE VALUE OF THE VECTOR IS A CHARACTER
class(seleno_thingy_clean[[1]])
## [1] "character"
is(seleno_thingy_clean[[1]])
## [1] "character" "vector"
## [3] "data.frameRowLabels" "SuperClassMethod"
## [5] "character_OR_connection" "character_OR_NULL"
## [7] "atomic" "EnumerationValue"
## [9] "vector_OR_Vector" "vector_OR_factor"
is.vector(seleno_thingy_clean[[1]])
## [1] TRUE
For old-times sake we can make a dotplot.
Now for a dotplot
WHAT AM I DOING HERE? TAKING A VECTOR OF THE LIST AND PUTTING INTO A SEPERATE OBJECT
prot1_vector <- seleno_thingy_clean[[1]]
prot2_vector <- seleno_thingy_clean[[2]]
We can dotplot like this
seqinr::dotPlot(prot1_vector,
prot1_vector)
WHAT DID I DO DIFFERENTLY HERE? YOU DID NOT TAKE A VECTOR OF THE LIST AND THE PLOT IS EVERY INDIVIDUAL CHARACTER
seqinr::dotPlot(seleno_thingy_clean[[1]],
seleno_thingy_clean[[2]])
dotPlot likes things in a single vector, but pairwiseAlignment like a single string of characters, so as always we have to process the data.
WHAT AM I DOING HERE? WHAT DOES "" MEAN? TAKING THE PASTE COMMAND AND SQUISHING EVERYTHING TOGETHER, THE ’’ ARE THE SPACES IN BETWEEN THAT WE ARE GETTING RID OF
prot1_str <- paste(seleno_thingy_clean[[1]],sep = "", collapse = "")
prot2_str <- paste(seleno_thingy_clean[[2]],sep = "", collapse = "")
So now things look like this HOW WOULD YOU DESCRIBE THIS? NOW THE VECTOR LENGTH = 1 WHERE EVERYTHING IS A LIST
prot1_str
## [1] "MERQEESLSARPALETEGLRFLHTTVGSLLATYGWYIVFSCILLYVVFQKLSARLRALRQRQLDRAAAAVEPDVVVKRQEALAAARLKMQEELNAQVEKHKEKLKQLEEEKRRQKIEMWDSMQEGKSYKGNAKKPQEEDSPGPSTSSVLKRKSDRKPLRGGGYNPLSGEGGGACSWRPGRRGPSSGGUG"
Protein alignments need a amino acid transition matrix, and we need to use data() to bring those up into active memory (VERY IMPORTANT STEP!)
data(BLOSUM50)
The alignment
align_out <- Biostrings::pairwiseAlignment(pattern = prot1_str,
subject = prot2_str,
type = "global",
gapOpening = -9.5,
gapExtension = -0.5)
What is this? THE RAW ALIGNMENT
align_out
## Global PairwiseAlignmentsSingleSubject (1 of 1)
## pattern: MERQEESLSARPALETEGLRFLHTTVGSLLATYG...-----------------ACSWRPGRRGPSSGGUG
## subject: M---------------------------------...IDIEPDIEALLSQGPSCA----------------
## score: -160.2561
WHAT IS THIS? HOW IS IT DIFFERNT FROM THE LAST CHUNK? THIS IS THE FULL ALIGNMENT WITH LOTS OF GAPS DUE TO THEM BEING TWO DIFFERENT SEQUENCES
compbio4all::print_pairwise_alignment(align_out)
## [1] "MERQEESLSARPALETEGLRFLHTTVGSLLATYGWYIVFSCILLYVVFQKLSARLRALRQ 60"
## [1] "M---------------------------------------C----------AARL----- 6"
## [1] " "
## [1] "RQLDRAAAAVEPDVVVKRQEALAAA--------RLKMQEELNAQVEKHKEKLKQLEEEKR 112"
## [1] "-----AAAA-------------AAAQSVYAFSAR-------------------------- 22"
## [1] " "
## [1] "RQKIEMWDSMQEGKSYKGNAKKPQEEDSPGPSTSSVLKRKSDRKPLRGGGYNPLSGE--- 169"
## [1] "--------------------------------------------PLAGG-------EPVS 31"
## [1] " "
## [1] "------------------------GGG--------------------------------- 172"
## [1] "LGSLRGKENAKNEEILNSLKYVRPGGGFEPNFMLFEKCEVNGAGAHPLFAFLREALPAPS 91"
## [1] " "
## [1] "------------------------------------------------------------ 172"
## [1] "DDATALMTDPKLITWSPVCRNDVAWNFEKFLVGPDGVPLRRYSRRFQTIDIEPDIEALLS 151"
## [1] " "
## [1] "-----A 227"
## [1] "QGPSCA 211"
## [1] " "
These are two randomly chosen sequences, so the alignment should be pretty …WHAT?
The score is negative, but on its own that MEANS WHAT? NO DEFINITIVE MEANING EVEN GOOD ALIGNMENTS CAN HAVE A NEGATIVE SCORE
score(align_out)
## [1] -160.2561
pid gives us … WHAT? BETTER SENSE OF THEY MATCH
pid(align_out)
## [1] 7.189542
Of course, pid can be calculated several ways (WHY IS THIS AN ISSUE / POSSIBLE?) IT IS AN ISSUE BECAUSE WE ARE GETTING SO MANY DIFFERENT VALUES
pid(align_out,type = "PID1")
## [1] 7.189542
pid(align_out,type = "PID2")
## [1] 91.66667
pid(align_out,type = "PID3")
## [1] 14.01274
pid(align_out,type = "PID4")
## [1] 12.71676