Detection of oxytetracycline production by Streptomyces rimosus in soil microcosms by combining whole-cell biosensors and flow cytometry

被引:54
作者
Hansen, LH
Ferrari, B
Sorensen, AH
Veal, D
Sorensen, SJ
机构
[1] Univ Copenhagen, Dept Gen Microbiol, DK-1307 Copenhagen K, Denmark
[2] Macquarie Univ, Dept Biol Sci, Sydney, NSW 2109, Australia
关键词
D O I
10.1128/AEM.67.1.239-244.2001
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Combining the high specificity of bacterial biosensors and the resolution power of fluorescence-activated cell sorting (FACS) provided qualitative detection of oxytetracycline production by Streptomyces rimosus in soil microcosms. A plasmid containing a transcriptional fusion between the tetR-regulated P-tet promoter from Tn10 and a FAGS-optimized gfp gene was constructed. When harbored by Escherichia coli, this plasmid produces large amounts of green fluorescent protein (GFP) in the presence of tetracycline. This tetracycline biosensor was used to detect the production of oxytetracycline by S. rimosus introduced into sterile soil. The tetracycline-induced GFP producing biosensors were detected by FAGS analysis, enabling the detection of oxytetracycline encounters by single biosensor cells. This approach can be used to study interactions between antibiotic producers and their target organisms in soil.
引用
收藏
页码:239 / 244
页数:6
相关论文
共 19 条
[1]   CONSTRUCTION OF A SINGLE-COPY PROMOTER VECTOR AND ITS USE IN ANALYSIS OF REGULATION OF THE TRANSPOSON-TN10 TETRACYCLINE RESISTANCE DETERMINANT [J].
BERTRAND, KP ;
POSTLE, K ;
WRAY, LV ;
REZNIKOFF, WS .
JOURNAL OF BACTERIOLOGY, 1984, 158 (03) :910-919
[2]   Phenotypic diversity and antibiotic resistance in soil bacterial communities [J].
Bronstad, K ;
Dronen, K ;
Ovreas, L ;
Torsvik, V .
JOURNAL OF INDUSTRIAL MICROBIOLOGY, 1996, 17 (3-4) :253-259
[3]   FACS-optimized mutants of the green fluorescent protein (GFP) [J].
Cormack, BP ;
Valdivia, RH ;
Falkow, S .
GENE, 1996, 173 (01) :33-38
[4]   TERRAMYCIN, A NEW ANTIBIOTIC [J].
FINLAY, AC ;
HOBBY, GL ;
PAN, SY ;
REGNA, PP ;
ROUTIEN, JB ;
SEELEY, DB ;
SHULL, GM ;
SOBIN, BA ;
SOLOMONS, IA ;
VINSON, JW ;
KANE, JH .
SCIENCE, 1950, 111 (2874) :85-85
[5]   PRODUCTION AND ROLE OF ANTIBIOTICS IN SOIL [J].
GOTTLIEB, D .
JOURNAL OF ANTIBIOTICS, 1976, 29 (10) :988-1000
[6]   STUDIES ON TRANSFORMATION OF ESCHERICHIA-COLI WITH PLASMIDS [J].
HANAHAN, D .
JOURNAL OF MOLECULAR BIOLOGY, 1983, 166 (04) :557-580
[7]   Detection and quantification of tetracyclines by whole cell biosensors [J].
Hansen, LH ;
Sorensen, SJ .
FEMS MICROBIOLOGY LETTERS, 2000, 190 (02) :273-278
[8]  
Hopwood D.A., 1985, GENETIC MANIPULATION
[9]  
Normander B, 1998, APPL ENVIRON MICROB, V64, P1902
[10]   Application of a mer-lux biosensor for estimating bioavailable mercury in soil [J].
Rasmussen, LD ;
Sorensen, SJ ;
Turner, RR ;
Barkay, T .
SOIL BIOLOGY & BIOCHEMISTRY, 2000, 32 (05) :639-646