Team:Bielefeld-Germany/Results/tvel5

TVEL5 integrated in shuttle vector

The laccase Trametes versicolor TVEL5 [http://partsregistry.org/wiki/index.php?title=Part:BBa_K863030 BBa_K863030] was cloned with classic restriction ligation steps into the shuttle vector ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K863204 BBa_K863204]). This construct ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K863207 BBa_K863207]) was linearized and integrated into the yeast genome of P. pastoris GS115. Positive Clones with single cross over were identified by PCR as shown in the following pictures. <<<<<<<BILDER PCR (2 pieces)>>>>>>>> The clones were cultivated in minimal medium and induced with 1 % (v/v) methanol twice a day for 2 days. The supernatant of the culture was examined to active laccases by testing the enzyme activity.

Figure 1: Activity test of cultivation (culture 1, 2, 3) supernatant of [http://partsregistry.org/Part:BBa_K863207 BBa_K863207]. The control is the P. pastoris GS115.