Fig. 5. PCR amplification and cloning of the putative ßARK homologue in Paramecium. (A) PCR product of 400 bp (lane 2, arrowhead) was electrophoretically transferred onto DEAE-cellulose. DNA was eluted and used as a template for PCR reamplification resulting in the product of predicted molecular size of 400 bp (B; lane 2, arrowhead). Following subcloning into pGEM-T vector and transfection, the plasmid DNA was isolated from the positive colonies (as described in Materials and methods) and subsequently digested with EcoRI. The presence of an insert of the correct size was detected (C; lane 2, arrowhead). Lanes 1 in A–C are molecular mass standards.