Molecular cloning in Escherichia coli of Erwinia chrysanthemi genes encoding multiple forms of pectate lyase
10.1128/jb.161.3.913-920.1985
Crossref journal-article
American Society for Microbiology
Journal of Bacteriology (235)
Abstract

The phytopathogenic enterobacterium Erwinia chrysanthemi excretes multiple isozymes of the plant tissue-disintegrating enzyme, pectate lyase (PL). Genes encoding PL were cloned from E. chrysanthemi CUCPB 1237 into Escherichia coli HB101 by inserting Sau3A-generated DNA fragments into the BamHI site of pBR322 and then screening recombinant transformants for the ability to sink into pectate semisolid agar. Restriction mapping of the cloned DNA in eight pectolytic transformants revealed overlapping portions of a 9.8-kilobase region of the E. chrysanthemi genome. Deletion derivatives of these plasmids were used to localize the pectolytic genotype to a 2.5-kilobase region of the cloned DNA. PL gene expression in E. coli was independent of vector promoters, repressed by glucose, and not induced by galacturonan. PL accumulated largely in the periplasmic space of E. coli. An activity stain used in conjunction with ultrathin-layer isoelectric focusing resolved the PL in E. chrysanthemi culture supernatants and shock fluids of E. coli clones into multiple forms. One isozyme with an apparent pI of 7.8 was produced at a far higher level in E. coli and was common to all of the pectolytic clones. Activity staining of renatured PL in sodium dodecyl sulfate-polyacrylamide gels revealed that this isozyme comigrated with the corresponding isozyme produced by E. chrysanthemi. The PL isozyme profiles produced by different clones and deletion derivative subclones suggest that the cloned region contains at least two PL isozyme structural genes. Pectolytic E. coli clones possessed a limited ability to macerate potato tuber tissues.

Bibliography

Collmer, A., Schoedel, C., Roeder, D. L., Ried, J. L., & Rissler, J. F. (1985). Molecular cloning in Escherichia coli of Erwinia chrysanthemi genes encoding multiple forms of pectate lyase. Journal of Bacteriology, 161(3), 913–920.

Authors 5
  1. A Collmer (first)
  2. C Schoedel (additional)
  3. D L Roeder (additional)
  4. J L Ried (additional)
  5. J F Rissler (additional)
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Dates
Type When
Created 5 years, 7 months ago (Jan. 3, 2020, 1:05 p.m.)
Deposited 4 years, 1 month ago (July 29, 2021, 1:42 p.m.)
Indexed 1 year, 10 months ago (Oct. 25, 2023, 1:42 a.m.)
Issued 40 years, 6 months ago (March 1, 1985)
Published 40 years, 6 months ago (March 1, 1985)
Published Print 40 years, 6 months ago (March 1, 1985)
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@article{Collmer_1985, title={Molecular cloning in Escherichia coli of Erwinia chrysanthemi genes encoding multiple forms of pectate lyase}, volume={161}, ISSN={1098-5530}, url={http://dx.doi.org/10.1128/jb.161.3.913-920.1985}, DOI={10.1128/jb.161.3.913-920.1985}, number={3}, journal={Journal of Bacteriology}, publisher={American Society for Microbiology}, author={Collmer, A and Schoedel, C and Roeder, D L and Ried, J L and Rissler, J F}, year={1985}, month=mar, pages={913–920} }