dist_stopcodon <- function(target_peakcenter,annotation_file){
##get the stop codon site
txdbfile <- GenomicFeatures::makeTxDbFromGFF(annotation_file)
genes_txdb <- genes(txdbfile)
exbytx_txdb <- exonsBy(txdbfile,by = "tx")
isoform_ambiguity_method = "longest_tx"
if(isoform_ambiguity_method == "longest_tx"){
Longest_tx_table <- find_longest_transcript(exbytx_txdb,txdbfile)
Kept_tx_indx <- Longest_tx_table$TXID[Longest_tx_table$longest]
rm(Longest_tx_table)
} else {
Kept_tx_indx <- T
}
exbytx_txdb <- exbytx_txdb[Kept_tx_indx]
exbytx_txdb <- exbytx_txdb[countOverlaps(exbytx_txdb,exbytx_txdb) == 1]
txid <- names(exbytx_txdb)
cds <- cdsBy(txdbfile,by = "tx")
cds <- cds[ names(cds) %in% txid ]
Stop_codons <- resize( unlist( range(cds) ) , 3, fix = "end" )
Stopcodon <- resize(Stop_codons,1,fix = "center")
##peak center
target_sites_se <- GRanges(seqnames = as.character(target_peakcenter$seqnames),
IRanges(start = as.numeric(as.character(target_peakcenter$start)),
width = 1),strand = as.character(target_peakcenter$strand))
target_sites_map <- mapToTranscripts(target_sites_se, exbytx_txdb,ignore.strand=F)
stopcodon_map <- mapToTranscripts(Stopcodon,exbytx_txdb)
##
overlap_hit_tx <- intersect(as.character(seqnames(target_sites_map)),
as.character(seqnames(stopcodon_map)))
select_targetsites_map <- target_sites_map[which(!is.na(match(as.character(seqnames(target_sites_map)),overlap_hit_tx)))]
select_peaknum <- as.numeric(as.character(select_targetsites_map$xHits))
fun_hit <- function(overlap_hit){
site_label <- which(!is.na(match(as.character(seqnames(select_targetsites_map)),overlap_hit)))
stopcodon_label <- which(!is.na(match(as.character(seqnames(stopcodon_map)),overlap_hit)))
site_pos <- start(select_targetsites_map[site_label])
stopcodon_pos <- start(stopcodon_map[stopcodon_label])
# tx_width <- as.numeric(sum(width(exbytx_txdb[which(!is.na(match(names(exbytx_txdb),overlap_hit)))])))
abs_pos <- abs(site_pos-stopcodon_pos)
# norm_pos <- abs_pos/tx_width
site_num <- select_targetsites_map[site_label]$xHits
names(abs_pos) <- site_num
return(abs_pos)
}
site_norm_pos <- mapply(fun_hit, overlap_hit_tx)
names(site_norm_pos) <- NULL
m6Asite_norm_pos <- unlist(site_norm_pos)
m6Asite_num <- as.numeric(as.character(names(m6Asite_norm_pos)))
select_m6Asite_overlap <- target_peakcenter[m6Asite_num,]
add_norm_pos_ifor <- data.frame(select_m6Asite_overlap,as.numeric(as.character(m6Asite_norm_pos)))
colnames(add_norm_pos_ifor)[ncol(add_norm_pos_ifor)] <- "dist_stopcodon"
return(add_norm_pos_ifor)
}
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