Open AccessConstruction of a bioinsecticidal rhizosphere isolate of Pseudomonas fluorescens
Author(s) -
Waalwijk Cees,
Dullemans Annette,
Maat Corien
Publication year - 1991
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1991.tb04358.x
Subject(s) - pseudomonas fluorescens , biology , pseudomonas , bacillus thuringiensis , plasmid , southern blot , kanamycin , microbiology and biotechnology , anopheles stephensi , gene , molecular cloning , genetics , bacteria , gene expression , botany , aedes aegypti , larva
Summary The cryIVB gene from Bacillus thuringiensis morrisoni PG‐14 was cloned and expressed in Escherichia coli . A gene cassette was constructed that placed the gene under the control of the tac promotor. Three Pseudomonas ‘suicide’ vectors were made by cloning chromosomal DNA fragments from the root‐colonizing Pseudomonas fluorescens strain P1 into plasmid pSUP202. The kanamycin resistance gene nptII and the cryIVB gene cassette were cloned within the Pseudomonas sequences. These constructs were introduced into the root‐colonizing strain Pseudomonas fluorescens P1. Southern blot hybridizations demonstrated that the nptII and cryIVB genes were integrated into the chromosome whereas vector sequences were not. Expression of the cryIVB protein by transgenic Pseudomonas cells was demonstrated by Western blot analysis. Cell cultures of the transformed P. fluorescens were found to be toxic towards larvae of the malaria mosquito Anopheles stephensi and to leatherjackets, the larvae of Tipula oleracea .