Use of Tn5tac1 to Clone a Pel Gene Encoding a Highly Alkaline, Asparagine-rich Pectate Lyase Isozyme from an Erwinia Chrysanthemi EC16 Mutant with Deletions Affecting the Major Pectate Lyase Isozymes

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1995 Aug 1

PMID 7635842

Citations 10

Authors

J. R. Alfano

J H Ham

A Collmer

Affiliations

Soon will be listed here.

Abstract

Erwinia chrysanthemi mutant CUCPB5047, delta(pelA pelE) delta(pelB pelC)::28bp delta(pelX) delta 4bp pehX::omega Cmr, was constructed, mutated with Tn5tac1, and screened for isopropyl-beta-D-thiogalactopyranoside-dependent pectate lyase (Pel) production. A Kmr SacI fragment from the hyperexpressing Pel+ mutant CUCPB5066 was cloned into Escherichia coli and sequenced. The gene identified, pelL, encodes a novel, asparagine-rich, highly alkaline enzyme that is similar in primary structure to PelX and in enzymological properties to PelE.

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