Mutant/mutation
The mutant (strongly) expresses GFP in blood stages (under the control of the constitutive and strong hsp70 promoter). This Bergreen mutant does contain the hdhfr/fcu drug-selectable marker. See RMgm-757 for a similar Bergreen mutant where the drug-selectable marker has been removed by negative selection. The gfp-gene has been integrated by double cross-over integration into a silent intergenic region on chromosome 6.

Phenotype
Blood stages, oocysts, sporozoites and liver stages (strongly) express GFP See RMgm-757 for characterization of GFP expression in all life cycle stages for an independent P. berghei Bergreen mutant).

Additional information
To generate the three isogenic, strongly fluorescent reference parasite lines, Bergreen, Beryellow (RMgm-948), and Berred (RMgm-949), that express high levels of GFP, YFP, and mCherry, respectively,  the mCherry-3xMyc carboxy-terminal tagging sequence was removed from the original pBAT-SIL6 vector (RMgm-757) by restriction digestion with BlpI and HpaI followed by Klenow fill-in and religation of the plasmid. The resulting vector pBAT-G6 harbors the GFP-expression and recyclable drug-selectable cassettes along with integration sequences targeting the silent, intergenic locus on P. berghei chromosome 6 (seeRMgm- 757 for details of the pBAT-SIL6 vector). The GFP from the high-expressing fluorescent protein cassette was exchanged for YFP and mCherry to yield pBAT-Y6 and pBAT-M6, respectively. The plasmids were verified by commercial Sanger sequencing and linearized with ApaLI and AhdI before transfection into wild-type P. berghei strain ANKA parasites.
Isolation of fluorescent parasites was performed by FACS-sorting, with the following adaptation to sort in three different channels. We used excitation wavelengths of 488 nm for GFP and YFP and 561 nm for mCherry parasites. Fluorescence was detected using the following band pass filters: GFP, 513/17 nm; YFP, 530/30nm; mCherry, 610/20 nm.

Other mutants