Nishida T, Suzuki T, Ito K, Kamio Y, Izaki K
Department of Agricultural Chemistry, Faculty of Agriculture, Tohoku University, Sendai, Japan.
Biochem Biophys Res Commun. 1990 Apr 30;168(2):801-8. doi: 10.1016/0006-291x(90)92392-d.
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 (0RF) 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.
