CHARACTERIZATION AND OVEREXPRESSION OF THE PEM GENE ENCODING PECTIN METHYLESTERASE OF ERWINIA-CHRYSANTHEMI STRAIN-3937

被引：35

作者：

LAURENT, F ^{[1
]}

KOTOUJANSKY, A ^{[1
]}

LABESSE, G ^{[1
]}

BERTHEAU, Y ^{[1
]}

机构：

[1] UNIV PARIS 06,UNIV PARIS 07,MINERAL CRISTALLOG LAB,CNRS,URA 09,F-75252 PARIS 05,FRANCE

来源：

GENE | 1993年 / 131卷 / 01期

关键词：

RECOMBINANT PLASMID; PROMOTER; SOFT ROT DISEASE; PECTINOLYTIC ENZYME; PECTIN;

D O I：

10.1016/0378-1119(93)90664-O

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

The pem gene encoding the pectin methylesterase (PME) of Erwinia chrysanthemi strain 3937 was subcloned and its nucleotide sequence determined. The gene contains an open reading frame of 1098 bp and codes for a protein of 366 amino acids (aa). The mature 37-kDa form of the protein is 342 aa long and has a calculated isoelectric point of 9.64. A plasmid was constructed to overproduce PME: a DNA fragment carrying pem was amplified by the polymerase chain reaction and cloned downstream from the p(L) promoter of the lambda phage, in a high-copy-number plasmid. In an Escherichia coli strain transformed with this plasmid, an increase in PME production of more than 60-fold was obtained, compared with the wild-type Er. chrysanthemi strain. PME represents about 5% of the total protein content of the cells. Comparison of this PME sequence with six PMEs from prokaryotic or eukaryotic organisms showed six highly conserved segments whose possible role in enzyme activity are discussed.

引用

页码：17 / 25

页数：9

共 45 条

[1] A GENE SHOWING SEQUENCE SIMILARITY TO PECTIN ESTERASE IS SPECIFICALLY EXPRESSED IN DEVELOPING POLLEN OF BRASSICA-NAPUS - SEQUENCES IN ITS 5' FLANKING REGION ARE CONSERVED IN OTHER POLLEN-SPECIFIC PROMOTERS [J].