CLONING AND SEQUENCE OF A 3-CENTER-DOT-835 KBP DNA FRAGMENT CONTAINING THE HIS4 GENE AND A FRAGMENT OF A PEX5-LIKE GENE FROM CANDIDA-ALBICANS

We have isolated the Candida albicans HIS4 (CaHIS4) gene by complement ation of a his4-34 Saccharomyces cerevisiae mutant. The sequenced DNA fragment contains a putative ORF of 2514 bp, whose translation product shares a global identity of 44% and 55% to the His4 protein homologs of S. cerevisiae and Kluyveromyces lactis, respectively. Analysis of C aHIS4 sequence suggests that, similarly to S. cerevisiae HIS4, it code s for a polypeptide having three separate enzymatic activities (phosph oribosyl-AMP cyclohydrolase, phosphoribosyl-ATP pyrophosphohydrolase a nd histidinol dehydrogenase) which reside in different domains of the protein. A C. albicans his4 strain is complemented with this gene when using a C, albicans-S. cerevisiae-Escherichia coli shuttle vector, th us enabling the construction of a host system for C. albicans genetic manipulation. In addition, upstream of the sequenced CaHIS4 sequence, we have found the 3'-terminal half of a gene encoding a PEX5-like prot ein. The EMBL/DDJB/GenBank Accession Number of this sequence is AJ0031 15. (C) 1998 John Wiley & Sons, Ltd.