CLONING AND EXPRESSION OF PECTIN LYASE GENE FROM ERWINIA-CAROTOVORA IN ESCHERICHIA-COLI

被引:8
|
作者
NISHIDA, T
SUZUKI, T
ITO, K
KAMIO, Y
IZAKI, K
机构
[1] Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University, Sendai
关键词
D O I
10.1016/0006-291X(90)92392-D
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A pectin lyase (PNL; EC 4.2.2.10) gene of Erwinia carotovora Er was cloned and expressed in Escherichia coli. The analysis of the nucleotide sequence of the 0.6 kb StuI-EcoRI fragment, which was hybridized with the mixed oligonucleotide probe for PNL gene, revealed the presence of an open reading frame (ORF) and correlated exactly with the known N-terminal 18 amino acid sequence of PNL. When a plasmid pTN2159, which has a BamHI-EcoRI fragment containing this ORF, was introduced into E. coli JM109, PNL was not expressed. When a tac-promoter was inserted in front of the ORF, PNL was efficiently expressed in E. coli. Synthesis of PNL by E. coli was also confirmed by immunoblot analysis. © 1990.
引用
收藏
页码:801 / 808
页数:8
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