Construction of chromosomally encoded secreted hemolysin fusion proteins by use of mini-TnhlyAs transposon.
We used the minitransposon TnhlyAs [Gentschev, I., Maier, G., Kranig, A. and Goebel, W. (1996) Mol. Gen. Genet. 252, 266-274] for random insertion of the secretion signal (HlyAs) of Escherichia coli hemolysin (HlyA) into chromosomal genes. Four mini-TnhlyAs derivatives bearing the gltA (citrate synthase), deoC (2 deoxyribose-5 phosphate aldolase), tig (trigger factor) genes and an unknown ORF fused to hlyAs were identified and characterized. Our data suggest that TnhlyAs-generated hemolysin fusion proteins are secreted efficiently by the HlyB/HlyD/TolC hemolysin secretion machinery and that this can be useful for studies of gene expression or function.
Lehrstuhl für Mikrobiologie, Theodor-Boveri-Institut für Biowissenschaften, Würzburg, Germany. gentsch@biozentrum.uni-wuerzburg.de
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Gentschev
D001483:Base Sequence; D002950:Citrate (si)-Synthase / Q000235:genetics; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D006460:Hemolysin Proteins / Q000096:biosynthesis / Q000235:genetics*; D015151:Immunoblotting; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics* / Q000378:metabolism; D017385:Sequence Homology
D004251:DNA Transposable Elements; D006460:Hemolysin Proteins; D011993:Recombinant Fusion Proteins; D002950:Citrate (si)-Synthase