R: "Unary operator error" from multiline ggplot2 command

This is a well-known nuisance when posting multiline commands in R. (You can get different behavior when you source() a script to when you copy-and-paste the lines, both with multiline and comments)

Rule: always put the dangling '+' at the end of a line so R knows the command is unfinished:

ggplot(...) + geom_whatever1(...) +
  geom_whatever2(...) +
  stat_whatever3(...) +
  geom_title(...) + scale_y_log10(...)

Don't put the dangling '+' at the start of the line, since that tickles the error:

Error in "+ geom_whatever2(...) invalid argument to unary operator"

And obviously don't put dangling '+' at both end and start since that's a syntax error.

So, learn a habit of being consistent: always put '+' at end-of-line.

cf. answer to "Split code over multiple lines in an R script"

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.