1.Mutation of lambdapL/pR-cI857 system for production of bacterial ghost in Escherichia coli.
Hongliang DONG ; Xian'gan HAN ; Hao BAI ; Liang HE ; Lei LIU ; Rui LIU ; Tongjie CHAI ; Chan DING ; Haiwen LIU ; Shengqing YU
Chinese Journal of Biotechnology 2012;28(12):1423-1430
Bacterial ghost is intact envelope of Gram-negative bacteria, which is produced by the function of the lysis gene E from bacteriophage PhiX174. The expression of the lysis gene E is usually controlled by the thermosensitive lambdapL/pR-cI857 promoter. In this study, we described a mutation (T --> C) at the ninth nucleotide of the OR2 in the lambdapR promoter of the lambdapL/pR-cI857 system by overlap PCR. The bacteriolytic assay showed that the mutation in the lambdapL/pR-cI857 system enhanced the temperature of repressing the expression of gene E up to 37 degrees C. The lysis efficiency of altered lambdapR promoter in Escherichia coli DH5a and avian pathogenic E. coli DE17 was up to 99.9%. The expanded range of temperature will benefit for the production of bacterial ghost.
Bacteriolysis
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physiology
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Bacteriophage lambda
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genetics
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Base Sequence
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Cell Membrane
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physiology
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DNA, Bacterial
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analysis
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Escherichia coli
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genetics
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growth & development
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physiology
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virology
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Gene Expression Regulation
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genetics
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Molecular Sequence Data
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Mutation
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Promoter Regions, Genetic
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genetics
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Viral Proteins
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genetics
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metabolism