#' GO Enrichment analysis function
#' @importFrom dplyr filter_
#' @param df DGE files (DESeq2 result files) or vector contains gene names
#' @param GO_FILE GO annotation data
#' @param OP use BP,CC or MF
#' @param minSize minimal number of genes included in significant terms
#' @param padj.method p value adjust method (default: BH)
#' @param cutoff cutoff value for filtering significant terms (default: 0.05)
#' @param maxSize maximum number of genes included in significant terms
#' @param keepRich keep terms with rich factor value equal 1 or not (default: TRUE)
#' @param filename output filename
#' @param gene.cutoff the cut-off value for select DEGs (default: 0.01)
#' @export
#' @author Kai Guo
GE<-function(df,GO_FILE,OP="BP",gene.cutoff=0.01,minSize=2,maxSize=500,keepRich=TRUE,filename=NULL,padj.method="BH",cutoff=0.05){
go2gene<-sf(GO_FILE)
all_go<-.get_go_dat(ont = OP)
go2gene<-go2gene[names(go2gene)%in%rownames(all_go)];
gene2go<-reverseList(go2gene)
if(is.data.frame(df)){
IGE<-rownames(df)[df$padj<gene.cutoff]
}else{
IGE=as.vector(df)
}
fgene2go<-gene2go[IGE];
fgo2gene<-reverseList(fgene2go)
k=name_table(fgo2gene);
n=sum(!is.na(names(fgene2go)))
IGO<-names(fgo2gene);
N=length(unique(unlist(go2gene)));
M<-name_table(go2gene[IGO])
rhs<-hyper_bench_vector(k,M,N,n)
lhs<-p.adjust(rhs,method=padj.method)
rhs_an<-all_go[names(rhs),]
rhs_gene<-unlist(lapply(fgo2gene, function(x)paste(unique(x),sep="",collapse = ",")))
resultFis<-data.frame("Annot"=names(rhs),"Term"=rhs_an,"Annotated"=M[names(rhs)],
"Significant"=k[names(rhs)],"Pvalue"=as.vector(rhs),"Padj"=lhs,
"GeneID"=rhs_gene[as.vector(names(rhs))])
resultFis<-resultFis[order(resultFis$Pvalue),]
resultFis<-resultFis[resultFis$Pvalue<cutoff,]
resultFis<-filter_(resultFis, ~Significant<=maxSize)
if(keepRich==FALSE){
resultFis<-filter_(resultFis, ~Significant>=minSize)
}else{
resultFis<-filter_(resultFis, ~Significant>=minSize|(~Annotated/~Significant)==1)
}
if(!is.null(filename)){
write.table(resultFis,file=paste(filename,OP,"res.txt",sep="_"),sep="\t",quote=F,row.names=F)
}
return(resultFis);
}
#' Display GO enrichment result
#' @param resultFis: GO ennrichment analysis result data.frame
#' @param top: Number of Terms you want to display
#' @param filename: output filename
#' @param pvalue.cutoff: the cut-off value for selecting Term
#' @param padj.cutoff: the padj cut-off value for selecting Term
#' @param usePadj use adjust pvalue or not
#' @param low color used for small value
#' @param high color used for large value
#' @param order order bar or not
#' @param horiz use horiz or not
#' @param fontsize.x fontsize for x axis
#' @param fontsize.y fontsize for y axis
#' @param filename output filename
#' @param width width for output file
#' @param height height for output file
#' @param angle angle for x ticks label
#' @param horiz horizontal plot or not (default: FALSE)
#' @export
#' @author Kai Guo
GE.plot<-function(resultFis,top=50,pvalue.cutoff=0.05,order=FALSE,horiz=FALSE,
low="lightpink",high="red",
font.x="bold",font.y="bold",fontsize.x=10,fontsize.y=10,
fontsize.text=3,angle=75,padj.cutoff=NULL,usePadj=TRUE,
filename=NULL,width=10,height=8){
require(ggplot2)
if(!is.null(padj.cutoff)){
resultFis<-resultFis[resultFis$Padj<padj.cutoff,]
}else{
resultFis<-resultFis[resultFis$Pvalue<pvalue.cutoff,]
}
if(nrow(resultFis)>=top){
resultFis<-resultFis[1:top,]
}
if(max(resultFis$Significant/(resultFis$Annotated+0.1))<=1){
yheight=max(resultFis$Significant/resultFis$Annotated)+0.1
}else{
yheight=1
}
if(order==TRUE){
resultFis$rich<-as.numeric(resultFis$Significant)/as.numeric(resultFis$Annotated)
resultFis$Term<-factor(resultFis$Term,levels=resultFis$Term[order(resultFis$rich)])
}
if(usePadj==FALSE){
p<-ggplot(resultFis,aes(x=Term,y=round(as.numeric(Significant/Annotated),2)))+geom_bar(stat="identity",aes(fill=-log10(as.numeric(Pvalue))))
p<-p+scale_fill_gradient(low=low,high=high)+theme_light()
if(horiz==TRUE){
p<-p+theme(axis.text.y=element_text(face=font.y,size=fontsize.y),axis.text.x=element_text(face=font.x,color="black",size=fontsize.x,angle=angle))+labs(fill="-log10(Pvalue)")
p<-p+coord_flip()
p<-p+geom_text(aes(label=Significant),hjust=-0.3,size=fontsize.text)+xlab("Annotation")+ylab("Rich Factor")+ylim(0,yheight)
}else{
p<-p+theme(axis.text.y=element_text(face=font.y,size=fontsize.y),axis.text.x=element_text(face=font.x,color="black",size=fontsize.x,angle=angle,vjust=1,hjust=1))+labs(fill="-log10(Pvalue)")
p<-p+geom_text(aes(label=Significant),vjust=-0.3,size=fontsize.text)+xlab("Annotation")+ylab("Rich Factor")+ylim(0,yheight)
}
print(p)
}else{
p<-ggplot(resultFis,aes(x=Term,y=round(as.numeric(Significant/Annotated),2)))+geom_bar(stat="identity",aes(fill=-log10(as.numeric(Padj))))
p<-p+scale_fill_gradient2(low=low,high=high)+theme_light()
if(horiz==TRUE){
p<-p+theme(axis.text.y=element_text(face=font.y,size=fontsize.y),axis.text.x=element_text(face=font.x,color="black",size=fontsize.x,angle=angle))+labs(fill="-log10(Padj)")
p<-p+coord_flip()
p<-p+geom_text(aes(label=Significant),hjust=-0.3,size=fontsize.text)+xlab("Annotation")+ylab("Rich Factor")+ylim(0,yheight)
}else{
p<-p+theme(axis.text.y=element_text(face=font.y,size=fontsize.y),axis.text.x=element_text(face=font.x,color="black",size=fontsize.x,angle=angle,vjust=1,hjust=1))+labs(fill="-log10(Padj)")
p<-p+geom_text(aes(label=Significant),vjust=-0.3,size=fontsize.text)+xlab("Annotation")+ylab("Rich Factor")+ylim(0,yheight)
}
print(p)
}
if(!is.null(filename)){
ggsave(p,file=paste(filename,OP,"enrich.pdf",sep="_"),width=width,height=height)
}
}
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