Objective To study the genes involved in vacuole biogenesis in Candida albicans, so as to identify potential targets for antifungal drugs. Methods Five genes with potential involvement in vacuole biogenesis, ORF19.2734, ORF19.6950, ORF19.6605, ORF19.2006, and ORF19.5780, were selected for this study. The disruption of these target genes in Candida albicans was achieved by homologous recombination. Then the growth rate, hyphal formation, sensitivity to stresses, and the vacuole morphology of the mutants were analyzed. Results The disruptions of ORF19.6950, ORF19.6605, ORF19.2006, or ORF19.5780 resulted in no difference from the wild type strain in terms of growth rate, hyphal formation, sensitivity to stresses, and the vacuole morphology. The disruption of ORF19.2734 led to slower growth and higher sensitivity to ketoconazole, but showed no noticeable changes in vacuole mophogenesis. RT-PCR analysis found that mRNA expression of MDR1, ERG11, and ERG2 in the null mutant of ORF19.2734 was decreased compared with the wild type strain. Conclusion The null mutant of ORF19.2734 down-regulates the expression of MDR1, ERG11, and ERG2, therefore contributing to the increased sensitivity to azoles. Our results suggest that ORF19.2734 may be a potential treatment target for drug resistance in C. albicans.