Figure 1. Method for functional
complementation under yeast promoter control. Amplification of the
Upstream and Downstream Regulatory Regions (URR and DRR) is followed
by their ligation. This ligation product is amplified using the yORF-F
and yORF-R primers and inserted in pRS416 by BamHI ligation.
The human Open reading frame (BC002639) can subsequently be cloned
in the Bg/II site between the regulatory regions. Thus, extraneous
sequences are limited to 6 bp in the downstream region and 2 codons
at the 5'
of the ORF.