High level expression of Acinetobacter calcoaceticus mutarotase in Escherichia coli is achieved by improving the translational control sequence and removal of the signal sequence

The expression of enzymatically active Acinetobacter calcoaceticus encoded mutarotase in Escherichia coli is increased 10-fold upon fusion of the mro reading frame to an efficient ribosome binding sequence and deletion of the signal sequence. This was demonstrated by fusing the mro gene to the highl...

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Bibliographic Details
Published in:Applied microbiology and biotechnology 1989-05, Vol.30 (5), p.509-514
Main Authors: SCHMUCKER, R, GULLAND, U, WILL, M, HILLEN, W
Format: Article
Language:English
Subjects:
Acinetobacter calcoaceticus
Biological and medical sciences
Biotechnology
Fundamental and applied biological sciences. Psychology
Gene expression
gene fusion
Genetic technics
Methods. Procedures. Technologies
Molecular and cellular biology
Molecular genetics
Mutant screening
mutarotase
Prokaryotes
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Summary:The expression of enzymatically active Acinetobacter calcoaceticus encoded mutarotase in Escherichia coli is increased 10-fold upon fusion of the mro reading frame to an efficient ribosome binding sequence and deletion of the signal sequence. This was demonstrated by fusing the mro gene to the highly expressed tetR gene under control of the strong phage lambda promoter P sub(L). Deleting the leader sequence yielded cytoplasmic expression of 5 x 10 super(4) u of active mutarotase while constructions bearing the leader sequence expressed only 2 x 10 super(3) u active enzyme. It was shown by protein and Western blot analysis that the amounts of protein expressed are nearly the same with and without signal sequence.
ISSN:0175-7598
1432-0614
DOI:10.1007/BF00263857