火山图|给你geneList,帮我标到火山图上
火山图(Volcano Plot)常用于展示基因表达差异的分布,横坐标常为Fold change(倍数),越偏离中心差异倍数越大;纵坐标为P value(P值),值越大差异越显著。得名原因也许是因为结果图像火山吧!
火山图只标示指定基因?这需求都遇到过吧。
一 载入R包,数据
library(ggplot2)
library(openxlsx)
library(dplyr)
#绘制火山图数据
data <- read.xlsx("火山图.xlsx", sheet = 1)
head(data) #查看数据,主要有P值,Fold change和基因ID即可。
二 绘制火山图(标示最显著的基因)
2.1 先根据阈值分出上调和下调基因;
data$change <- as.factor(ifelse(data$adj.P.Val < 0.01 & abs(data$logFC) > 1,
ifelse(data$logFC > 1,'UP','DOWN'),'NOT'))
2.2 标示差异显著的基因
data$sign <- ifelse(data$adj.P.Val < 0.001 & abs(data$logFC) > 2.5,data$GENE_SYMBOL,NA)head(data)
2.3 绘制火山图
ggplot(data = data, aes(x = logFC, y = -log10(adj.P.Val), color = change)) +
geom_point(alpha=0.8, size = 1) +
theme_bw(base_size = 15) +
theme(panel.grid.minor = element_blank(),panel.grid.major = element_blank()) +
geom_hline(yintercept=2 ,linetype=4) +
geom_vline(xintercept=c(-1,1) ,linetype=4 ) +
scale_color_manual(name = "", values = c("red", "green", "black"), limits = c("UP", "DOWN", "NOT")) +
geom_text(aes(label = sign), size = 3)
了解一下ggplot2绘图的方式,标示的基因就是各个基因的text,然后想办法将其赋予到一个 aes 中即可。
三 标示指定基因
和上面类似,将指定基因添加到绘制数据中即可。
3.1 读入含有geneList的文件
gene <- read.xlsx("火山图.xlsx", sheet = 2)
gene$geneList <- gene$gene
额外生成一列相同列是为了后面合并后还有一列存在,这一列用于标示基因。(方法有点笨)
3.2 合并火山图数据
data2 <- data %>%
left_join(gene,by = c("GENE_SYMBOL" = "gene"))
head(data2)
增加了geneList列,为了后面使用text的方式添加上基因。
3.3 标示文件中的指定基因
ggplot(data = data2, aes(x = logFC, y = -log10(adj.P.Val), color = change)) +
geom_point(alpha=0.8, size = 1) +
theme_bw(base_size = 15) +
theme(panel.grid.minor = element_blank(),panel.grid.major = element_blank()) +
geom_hline(yintercept=2 ,linetype=4) +
geom_vline(xintercept=c(-1,1) ,linetype=4 ) +
scale_color_manual(name = "", values = c("red", "green", "black"), limits = c("UP", "DOWN", "NOT")) +
geom_text(aes(label = geneList), size = 5,color = "blue")
3.4 ggrepel 解决重叠问题
如果目标标示基因太多会导致重叠,可使用ggrepal函数
library(ggrepel)
ggplot(data = data2, aes(x = logFC, y = -log10(adj.P.Val), color = change)) +
geom_point(alpha=0.8, size = 1) +
theme_bw(base_size = 15) +
theme(panel.grid.minor = element_blank(),panel.grid.major = element_blank()) +
geom_hline(yintercept=2 ,linetype=4) +
geom_vline(xintercept=c(-1,1) ,linetype=4 ) +
scale_color_manual(name = "", values = c("red", "green", "black"), limits = c("UP", "DOWN", "NOT")) +
geom_label_repel(aes(label=geneList), fontface="bold", color="grey50", box.padding=unit(0.35, "lines"), point.padding=unit(0.5, "lines"), segment.colour = "grey50")
呐,可以随意标示感兴趣的基因了。