MmGFP6

Excerpts

We based MmGFP on mGFP5 (Siemering et al., 1996), which has two mutations, V163A and S175G, that improve the folding of the apoprotein and, therefore, its solubility, at higher temperatures. There are also codon usage changes that remove a cryptic splice site recognised in plants (Haseloff and Amos, 1995). Two further mutations, F64L and S65T, were introduced by PCR mutagenesis. The S65T mutation enhances the blue light absorption of GFP (Heim et al., 1995) and the F64L mutation (Cormack et al., 1996) enhances its solubility in bacterial cells, and therefore may also improve apoprotein folding in mammalian cells. (MmGFP also contains two apparently silent mutations, H25Y and G191D, that do not measurably affect fluorescence intensity.) MmGFP matures well at 37°C and is easily detectable in living mammalian cells using a standard FITC filter set.

Zernicka-Goetz et al. (1997)