Development of Streptococcus thermophilus lacZ as a reporter gene for Candida albicans

被引:66
作者
Uhl, MA [1 ]
Johnson, AD [1 ]
机构
[1] Univ Calif San Francisco, Dept Microbiol & Immunol, San Francisco, CA 94143 USA
来源
MICROBIOLOGY-SGM | 2001年 / 147卷
关键词
beta-galactosidase; yeast; fungal pathogen; gene regulation;
D O I
10.1099/00221287-147-5-1189
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The study of gene regulation in many organisms has been facilitated by the development of reporter genes. The authors report the use of lacZ from Streptococcus thermophilus, a gene encoding a beta -galactosidase, as a reporter for the fungal pathogen Candida albicans. As test cases, Strep. thermophilus lacZ was placed under control of three different C. albicans promoters: MAL2 (maltase), inducible by maltose; HWP1 (hyphal cell wall protein), induced by conditions that promote filamentous growth; and ACT1 (actin). These constructs were each integrated into the C. albicans genome and beta -galactosidase activity was readily detected from these strains, but only under the appropriate growth conditions. beta -Galactosidase activity could be detected by several methods: quantitative liquid assays using permeabilized cells. colorimetric assays of colonies replicated to paper filters, and in situ coloration of colonies growing on medium containing the indicator X-Gal. These results show the usefulness of Strep. thermophilus lacZ as a monitor of gene regulation in this medically important yeast.
引用
收藏
页码:1189 / 1195
页数:7
相关论文
共 45 条
  • [1] COS1, a two-component histidine kinase that is involved in hyphal development in the opportunistic pathogen Candida albicans
    Alex, LA
    Korch, C
    Selitrennikoff, CP
    Simon, MI
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (12) : 7069 - 7073
  • [2] Ausubel F.M., 1992, CURRENT PROTOCOLS MO
  • [3] NUCLEOTIDE-SEQUENCE OF A REGULATORY REGION OF THE UIDA GENE IN ESCHERICHIA-COLI-K12
    BLANCO, C
    RITZENTHALER, P
    MATAGILSINGER, M
    [J]. MOLECULAR & GENERAL GENETICS, 1985, 199 (01): : 101 - 105
  • [4] Control of filament formation in Candida albicans by the transcriptional repressor TUP1
    Braun, BR
    Johnson, AD
    [J]. SCIENCE, 1997, 277 (5322) : 105 - 109
  • [5] Regulatory networks controlling Candida albicans morphogenesis
    Brown, AJP
    Gow, NAR
    [J]. TRENDS IN MICROBIOLOGY, 1999, 7 (08) : 333 - 338
  • [6] Brown DH, 1996, MOL GEN GENET, V251, P75, DOI 10.1007/BF02174347
  • [7] LARGE-SCALE ANALYSIS OF GENE-EXPRESSION, PROTEIN LOCALIZATION, AND GENE DISRUPTION SACCHAROMYCES-CEREVISIAE
    BURNS, N
    GRIMWADE, B
    ROSSMACDONALD, PB
    CHOI, EY
    FINBERG, K
    ROEDER, GS
    SNYDER, M
    [J]. GENES & DEVELOPMENT, 1994, 8 (09) : 1087 - 1105
  • [8] Defective hyphal development and avirulence caused by a deletion of the SSK1 response regulator gene in Candida albicans
    Calera, JA
    Zhao, XJ
    Calderone, R
    [J]. INFECTION AND IMMUNITY, 2000, 68 (02) : 518 - 525
  • [9] Yeast-enhanced green fluorescent protein (yEGFP): A reporter of gene expression in Candida albicans
    Cormack, BP
    Bertram, G
    Egerton, M
    Gow, NAR
    Falkow, S
    Brown, AJP
    [J]. MICROBIOLOGY-UK, 1997, 143 : 303 - 311
  • [10] Derepressed hyphal growth and reduced virulence in a VH1 family-related protein phosphatase mutant of the human pathogen Candida albicans
    Csank, C
    Makris, C
    Meloche, S
    Schroppel, K
    Rollinghoff, M
    Dignard, D
    Thomas, DY
    Whiteway, M
    [J]. MOLECULAR BIOLOGY OF THE CELL, 1997, 8 (12) : 2539 - 2551