CHARACTERIZATION AND INFLUENCE ON VIRULENCE OF THE LEVANSUCRASE GENE FROM THE FIREBLIGHT PATHOGEN ERWINIA-AMYLOVORA

被引:120
作者
GEIER, G
GEIDER, K
机构
[1] Max-Planck-Institut für medizinische Forschung, D-69028 Heidelberg
关键词
D O I
10.1006/pmpp.1993.1029
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
Levan, a β-2,6-fructofuranan, produced by the fireblight pathogen Erwinia amylovora, is synthesized from sucrose by the extracellular enzyme levansucrase. Levansucrase mutants, created by Tn5 transposon mutagenesis, showed retarded development of necrotic symptoms on inoculated pear seedlings. By complementation experiments the levansucrase gene (lsc) was located in a 1·4 kb PstI-EcoRV chromosomal DNA fragment, isolated from a genomic library of E. amylovora. An open reading frame of 415 amino acids, promotor sequences and a ribosome binding site were identified by DNA sequencing. The cloned lsc gene of E. amylovora was expressed in E. coli from its own promotor. The enzyme accumulated in E. coli in the periplasm and caused an altered colony morphology on sucrose containing solid media. When analysed by SDS-PAGE and isoelectric focusing, the enzyme purified from E. coli was found to be identical with the protein isolated from E. amylovora. © 1993 Academic Press, Inc.
引用
收藏
页码:387 / 404
页数:18
相关论文
共 57 条
[11]   PURIFICATION AND PROPERTIES OF AN EXTRACELLULAR LEVANSUCRASE FROM ERWINIA-HERBICOLA NRRL B-1678 [J].
COTE, GL ;
IMAM, SH .
CARBOHYDRATE RESEARCH, 1989, 190 (02) :299-307
[12]  
DANIELS MJ, 1988, ANNU REV PHYTOPATHOL, V26, P285, DOI 10.1146/annurev.py.26.090188.001441
[13]  
Dedonder R, 1966, METHOD ENZYMOL, V8, P500
[14]   BROAD HOST RANGE DNA CLONING SYSTEM FOR GRAM-NEGATIVE BACTERIA - CONSTRUCTION OF A GENE BANK OF RHIZOBIUM-MELILOTI [J].
DITTA, G ;
STANFIELD, S ;
CORBIN, D ;
HELINSKI, DR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1980, 77 (12) :7347-7351
[15]  
FALKENSTEIN H, 1989, J GEN MICROBIOL, V135, P2643
[16]   NUCLEOTIDE-SEQUENCE OF AN ESCHERICHIA-COLI CHROMOSOMAL HEMOLYSIN [J].
FELMLEE, T ;
PELLETT, S ;
WELCH, RA .
JOURNAL OF BACTERIOLOGY, 1985, 163 (01) :94-105
[17]   KINETIC BEHAVIOUR OF CALF-INTESTINAL ALKALINE PHOSPHATASE WITH 4-METHYLUMBELLIFERYL PHOSPHATE [J].
FERNLEY, HN ;
WALKER, PG .
BIOCHEMICAL JOURNAL, 1965, 97 (01) :95-&
[18]   POSITIVE SELECTION PROCEDURE FOR ENTRAPMENT OF INSERTION-SEQUENCE ELEMENTS IN GRAM-NEGATIVE BACTERIA [J].
GAY, P ;
LECOQ, D ;
STEINMETZ, M ;
BERKELMAN, T ;
KADO, CI .
JOURNAL OF BACTERIOLOGY, 1985, 164 (02) :918-921
[19]   GENETIC-ANALYSIS OF AN MDR-LIKE EXPORT SYSTEM - THE SECRETION OF COLICIN-V [J].
GILSON, L ;
MAHANTY, HK ;
KOLTER, R .
EMBO JOURNAL, 1990, 9 (12) :3875-3884
[20]   SECRETION OF CYCLOLYSIN, THE CALMODULIN-SENSITIVE ADENYLATE-CYCLASE HEMOLYSIN BIFUNCTIONAL PROTEIN OF BORDETELLA-PERTUSSIS [J].
GLASER, P ;
SAKAMOTO, H ;
BELLALOU, J ;
ULLMANN, A ;
DANCHIN, A .
EMBO JOURNAL, 1988, 7 (12) :3997-4004