R: “Unary operator error” from multiline ggplot2 command

It looks like you might have inserted an extra + at the beginning of each line, which R is interpreting as a unary operator (like - interpreted as negation, rather than subtraction). I think what will work is

ggplot(combined.data, aes(x = region, y = expression, fill = species)) +
    geom_boxplot() +
    scale_fill_manual(values = c("yellow", "orange")) + 
    ggtitle("Expression comparisons for ACTB") + 
    theme(axis.text.x = element_text(angle=90, face="bold", colour="black"))

Perhaps you copy and pasted from the output of an R console? The console uses + at the start of the line when the input is incomplete.