Comments | The BTU2 locus was modified in the CU428 strain background by addition of a sequence encoding mCherry. The resulting beta-tubulin protein has a c-terminal mCherry tag. The protein is expressed at its own locus. There is a linked neo4 cassette that confers resistance to paromomycin (100 ug/ml) in the presence of cadmium chloride (2.5 ug/ml). These cells may be resistant to paromomycin 100 ugml or higher without added cadmium. The strain is believed to be assorted in the macronucleus but this has not confirmed using molecular means. If the expression level decreases these cells need to be grown at the increasing concentrations of paromomycin to accumulate the tagged allele. |