CLONING, NUCLEOTIDE-SEQUENCE AND EXPRESSION OF A HEMOLYSIN GENE OF CLOSTRIDIUM-SEPTICUM

被引:17
作者
IMAGAWA, T [1 ]
DOHI, Y [1 ]
HIGASHI, Y [1 ]
机构
[1] OSAKA UNIV,COLL BIOMED TECHNOL,TOYONAKA,OSAKA 560,JAPAN
关键词
CLOSTRIDIUM SEPTICUM; HEMOLYSIN; DNA SEQUENCE;
D O I
10.1111/j.1574-6968.1994.tb06781.x
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A genomic library of Clostridium septicum NCTC547 strain was made in Escherichia coli by means of lambda gt10. The DNA insert of a hemolysin-positive (Hly(+)) lambda-clone was transferred into pUC19. The resulting plasmid, pCS21, confers a Hly(+) phenotype on E. coli. Crude lysates of E. coli (pCS21) possessed a strong lytic activity on human erythrocytes and also a lethal effect on mice, characteristic of an alpha toxin. Nucleotide sequence analysis revealed that the insert DNA (5.2 kb) in pCS21 included at least one open reading frame of 1380 bp. The coding frame for hemolysin was predicted to be 1329 bp in size and to encode a protein of 49.8 kDa. It coincided with the molecular mass (48 kDa) of the alpha toxin secreted by C. septicum. Taken together, the data indicated that plasmid pCS21 indeed encoded an cu toxin gene of C. septicum.
引用
收藏
页码:287 / 292
页数:6
相关论文
共 19 条
[1]   PURIFICATION AND CHARACTERIZATION OF THE LETHAL TOXIN (ALPHA-TOXIN) OF CLOSTRIDIUM-SEPTICUM [J].
BALLARD, J ;
BRYANT, A ;
STEVENS, D ;
TWETEN, RK .
INFECTION AND IMMUNITY, 1992, 60 (03) :784-790
[2]   Parallelism in the lethal and hemolytic activity of the toxin of Clostridium septicum [J].
Bernheimer, AW .
JOURNAL OF EXPERIMENTAL MEDICINE, 1944, 80 (04) :309-320
[3]   STRUCTURAL FEATURES OF MULTIPLE NIFH-LIKE SEQUENCES AND VERY BIASED CODON USAGE IN NITROGENASE GENES OF CLOSTRIDIUM-PASTEURIANUM [J].
CHEN, KCK ;
CHEN, JS ;
JOHNSON, JL .
JOURNAL OF BACTERIOLOGY, 1986, 166 (01) :162-172
[4]   CHARACTERIZATION OF A BACTERIOCINOGENIC PLASMID FROM CLOSTRIDIUM-PERFRINGENS AND MOLECULAR GENETIC-ANALYSIS OF THE BACTERIOCIN-ENCODING GENE [J].
GARNIER, T ;
COLE, ST .
JOURNAL OF BACTERIOLOGY, 1986, 168 (03) :1189-1196
[5]   TOXIGENIC CLOSTRIDIA [J].
HATHEWAY, CL .
CLINICAL MICROBIOLOGY REVIEWS, 1990, 3 (01) :66-98
[6]   CLONING AND NUCLEOTIDE SEQUENCING OF THE CLOSTRIDIUM-PERFRINGENS EPSILON-TOXIN GENE AND ITS EXPRESSION IN ESCHERICHIA-COLI [J].
HUNTER, SEC ;
CLARKE, IN ;
KELLY, DC ;
TITBALL, RW .
INFECTION AND IMMUNITY, 1992, 60 (01) :102-110
[7]   HIGH-EFFICIENCY TRANSFORMATION OF ESCHERICHIA-COLI WITH PLASMIDS [J].
INOUE, H ;
NOJIMA, H ;
OKAYAMA, H .
GENE, 1990, 96 (01) :23-28
[8]   COMPARISON OF THE ALPHA-TOXIN GENES OF CLOSTRIDIUM-PERFRINGENS TYPE-A AND TYPE-C STRAINS - EVIDENCE FOR EXTRAGENIC REGULATION OF TRANSCRIPTION [J].
KATAYAMA, S ;
MATSUSHITA, O ;
MINAMI, J ;
MIZOBUCHI, S ;
OKABE, A .
INFECTION AND IMMUNITY, 1993, 61 (02) :457-463
[9]   NUCLEOTIDE-SEQUENCE OF THE STREPTOLYSIN-O (SLO) GENE - STRUCTURAL HOMOLOGIES BETWEEN SLO AND OTHER MEMBRANE-DAMAGING, THIOL-ACTIVATED TOXINS [J].
KEHOE, MA ;
MILLER, L ;
WALKER, JA ;
BOULNOIS, GJ .
INFECTION AND IMMUNITY, 1987, 55 (12) :3228-3232
[10]   PROCEDURE FOR ISOLATION OF DEOXYRIBONUCLEIC ACID FROM MICRO-ORGANISMS [J].
MARMUR, J .
JOURNAL OF MOLECULAR BIOLOGY, 1961, 3 (02) :208-&