CLONING AND CHARACTERIZATION OF A GENE REQUIRED FOR THE SECRETION OF EXTRACELLULAR ENZYMES ACROSS THE OUTER-MEMBRANE BY XANTHOMONAS-CAMPESTRIS PV CAMPESTRIS

被引:76
作者
HU, NT [1 ]
HUNG, MN [1 ]
CHIOU, SJ [1 ]
TANG, F [1 ]
CHIANG, DC [1 ]
HUANG, HY [1 ]
WU, CY [1 ]
机构
[1] NATL CHUNG HSING UNIV,GRAD INST BIOL,TAICHUNG 40227,TAIWAN
关键词
D O I
10.1128/jb.174.8.2679-2687.1992
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Nonpathogenic mutants of Xanthomonas campestris pv. campestris, generated from transposon mutagenesis, accumulated extracellular polygalacturonate lyase, alpha-amylase, and endoglucanase in the periplasm. The transposon Tn5 was introduced by a mobilizable, suicidal plasmid, pSUP2021 or pEYDG1. Genomic banks of wild-type X. campestris pv. campestris, constructed on the broad-host-range, mobilizable cosmid pLAFR1 or pLAFR3, were conjugated with one of the mutants, designated XC1708. Recombinant plasmids isolated by their ability to complement XC1708 can be classified into two categories. One, represented by pLASC3, can complement some mutants, whereas the other, represented by a single plasmid, pLAHH2, can complement all of the other mutants. Restriction mapping showed that the two recombinant plasmids shared an EcoRI fragment of 8.9 kb. Results from subcloning, deletion mapping, and mini-Mu insertional mutation of the 8.9-kb EcoRI fragment suggested that a 4.2-kb fragment was sufficient to complement the mutant XC1708. Sequence analysis of this 4.2-kb fragment revealed three consecutive open reading frames (ORFs), ORF1, ORF2, and ORF3. Hybridization experiments showed that Tn5 in the genome of XC1708 and other mutants complemented by pLASC3 was located in ORF3, which could code for a protein of 83.5 kDa. A signal peptidase II processing site was identified at the N terminus of the predicted amino acid sequence. Sequence homology of 51% was observed between the amino acid sequences predicted from ORF3 and the pulD gene of Klebsiella species.
引用
收藏
页码:2679 / 2687
页数:9
相关论文
共 51 条
[1]   MUTANTS OF ERWINIA-CHRYSANTHEMI DEFECTIVE IN SECRETION OF PECTINASE AND CELLULASE [J].
ANDRO, T ;
CHAMBOST, JP ;
KOTOUJANSKY, A ;
CATTANEO, J ;
BERTHEAU, Y ;
BARRAS, F ;
VANGIJSEGEM, F ;
COLENO, A .
JOURNAL OF BACTERIOLOGY, 1984, 160 (03) :1199-1203
[2]  
BERGMEYER HU, 1983, METHOD ENZYMAT AN, V3, P171
[3]   SECRETION AND MEMBRANE INTEGRATION OF A FILAMENTOUS PHAGE-ENCODED MORPHOGENETIC PROTEIN [J].
BRISSETTE, JL ;
RUSSEL, M .
JOURNAL OF MOLECULAR BIOLOGY, 1990, 211 (03) :565-580
[4]   PLASMID INSERTION MUTAGENESIS AND LAC GENE FUSION WITH MINI-MU BACTERIOPHAGE TRANSPOSONS [J].
CASTILHO, BA ;
OLFSON, P ;
CASADABAN, MJ .
JOURNAL OF BACTERIOLOGY, 1984, 158 (02) :488-495
[5]   UNUSUAL GENETIC PHENOMENA ASSOCIATED WITH TN5 MUTAGENESIS IN ALCALIGENES-EUTROPHUS STRAIN H-1 [J].
CHOW, WYW ;
PETERSON, JB ;
ATHERLY, AG .
ARCHIVES OF MICROBIOLOGY, 1989, 152 (03) :289-295
[6]  
CHU ST, 1981, CHINESE J MICROBIOL, V14, P156
[7]  
DANIELS MJ, 1984, J GEN MICROBIOL, V130, P2447
[8]  
DENFERT C, 1989, J BIOL CHEM, V264, P17462
[9]   CLONING AND EXPRESSION IN ESCHERICHIA-COLI OF THE KLEBSIELLA-PNEUMONIAE GENES FOR PRODUCTION, SURFACE LOCALIZATION AND SECRETION OF THE LIPOPROTEIN PULLULANASE [J].
DENFERT, C ;
RYTER, A ;
PUGSLEY, AP .
EMBO JOURNAL, 1987, 6 (11) :3531-3538
[10]   KLEBSIELLA-PNEUMONIAE PULS GENE ENCODES AN OUTER-MEMBRANE LIPOPROTEIN REQUIRED FOR PULLULANASE SECRETION [J].
DENFERT, C ;
PUGSLEY, AP .
JOURNAL OF BACTERIOLOGY, 1989, 171 (07) :3673-3679