Figure 2.

Deletion of NhKIN1 from the wild-type genome. The wild-type chromosome shows NhKIN1, the deleted region, and RAD6, an apparent homologue of genes encoding ubiquitin-conjugating proteins. The SphI-KpnI fragment of pTMS5 was used to delete most of NhKIN1. A double cross-over event resulted in the recombinant chromosome, which is deleted for a region starting 182 bp 5′ of the translational start site and stopping 26 bp 5′ of the translation stop site of NhKIN1. The deleted sequence was replaced with hygB. Certain restriction enzyme sites mentioned in the text are noted: N, NheI; Ap, ApaLI; K, KpnI; S, SacII; and Sp, SphI.