A new proteic killer gene system, hig, was identified on the plasmid Rts1. The hig locus consisting of higA and higB is directly related to the temperature sensitive host cell growth conferred by Rts1. We proved that higB encoding presumably a 92-amino-acid polypeptide inhibited segregation of plasmid free cells, and higA encoding a 104-amino-acid polypeptide suppressed the HigB function both in cis and in trans. (C) 1996 Academic Press, Inc.