THE INFLUENCE OF MESSENGER-RNA SECONDARY STRUCTURE ON EXPRESSION OF ENDOGLUCANASE IN BACILLUS-SUBTILIS

被引:3
作者
LEE, BR [1 ]
KIM, JH [1 ]
PACK, MY [1 ]
机构
[1] KAIST,DEPT BIOTECHNOL,TAEJON 305701,SOUTH KOREA
关键词
D O I
10.1007/BF01022392
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
A gene for endoglucanase of Bacillus subtilis has been inserted into a Bacillus expression plasmid containing a strong BJ27 promoter and a synthetic ribosome binding site. Secondary structure analysis of mRNA showed the presence of a strong hairpin loop burying the SD sequence and the initiation codon. Alteration of secondary structure at this site by deletion analysis revealed a correlation between endoglucanase expression and accessibility of the ribosome binding site. Elimination of secondary structures increased endoglucanase expression over five-fold to a level at which endoglucanase occupied 60% of total protein which was secreted into culture medium.
引用
收藏
页码:1001 / 1006
页数:6
相关论文
共 11 条
[1]  
BRADFORD P, 1983, ANAL BIOCHEM, V131, P333
[2]   POSTTRANSCRIPTIONAL MODIFICATION OF MESSENGER-RNA CONFORMATION - MECHANISM THAT REGULATES ERYTHROMYCIN-INDUCED RESISTANCE [J].
HORINOUCHI, S ;
WEISBLUM, B .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES, 1980, 77 (12) :7079-7083
[3]   CONSTRUCTION OF A BACILLUS-SUBTILIS DOUBLE MUTANT DEFICIENT IN EXTRACELLULAR ALKALINE AND NEUTRAL PROTEASES [J].
KAWAMURA, F ;
DOI, RH .
JOURNAL OF BACTERIOLOGY, 1984, 160 (01) :442-444
[4]   OVERPRODUCTION OF EXTRACELLULAR ENDOGLUCANASE BY GENETICALLY ENGINEERED BACILLUS-SUBTILIS [J].
KIM, JH ;
PACK, MY .
BIOTECHNOLOGY LETTERS, 1993, 15 (02) :133-138
[5]   UTILIZATION OF ESCHERICHIA-COLI TAC PROMOTER IN BACILLUS-SUBTILIS [J].
LEE, BR ;
PACK, MY .
BIOTECHNOLOGY LETTERS, 1990, 12 (12) :925-930
[6]  
Maniatis T, 1989, MOL CLONING
[7]   USE OF DINITROSALICYLIC ACID REAGENT FOR DETERMINATION OF REDUCING SUGAR [J].
MILLER, GL .
ANALYTICAL CHEMISTRY, 1959, 31 (03) :426-428
[8]   NUCLEOTIDE-SEQUENCES THAT SIGNAL THE INITIATION OF TRANSCRIPTION AND TRANSLATION IN BACILLUS-SUBTILIS [J].
MORAN, CP ;
LANG, N ;
LEGRICE, SFJ ;
LEE, G ;
STEPHENS, M ;
SONENSHEIN, AL ;
PERO, J ;
LOSICK, R .
MOLECULAR & GENERAL GENETICS, 1982, 186 (03) :339-346
[9]  
Shaw W V, 1975, Methods Enzymol, V43, P737