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231222s1998 xx ||||| 00| ||eng c |
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|a pubmed25n0327.xml
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|a (DE-627)NLM097994782
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|a (NLM)9863057
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|a DE-627
|b ger
|c DE-627
|e rakwb
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|a eng
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|a Kalabat, D Y
|e verfasserin
|4 aut
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|a Chitobiase, a new reporter enzyme
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|c 1998
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|a Text
|b txt
|2 rdacontent
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|a ohne Hilfsmittel zu benutzen
|b n
|2 rdamedia
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|a Band
|b nc
|2 rdacarrier
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|a Date Completed 11.03.1999
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|a Date Revised 28.09.2018
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|a published: Print
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|a Citation Status MEDLINE
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|a N,N'-diacetylchitobiase (chitobiase) from the marine organism Vibrio harveyi is a highly stable reporter enzyme for gene fusions. This enzyme hydrolyzes the disaccharide chitobiose to N-acetyl glucosamine. The advantages of the reporter gene encoding chitobiase (chb) are: (i) that chitobiase and N-acetyl-beta-D-glucosaminidase activities are missing in E. coli strains, (ii) chitobiase can be monitored using blue/white colony indicator plates and (iii) convenient substrates for this enzyme are commercially available. The use of chitobiase as a reporter enzyme is generally applicable to the study of gene expression in those bacteria that do not contain N-acetyl-beta-D-glucosaminidases. We constructed plasmid vectors containing a multiple cloning site for producing in-frame fusions to chitobiase, the attP of lambda phase for movement into the bacterial chromosome for single-copy analysis, the gene encoding chloramphenicol acetyltransferase (cat), the pACYC184 origin of replication and the rrnBt1t2 terminator region upstream of the chb gene to prevent read-through from other promoters. In-frame fusions between the dnaA gene and chb were moved to the chromosome by site-specific recombination with the chromosomal attB site. These single-copy fusions were assayed for chitobiase to examine the effects of a deletion in the dnaA regulatory region
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|a Journal Article
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|a Research Support, U.S. Gov't, Non-P.H.S.
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|a Research Support, U.S. Gov't, P.H.S.
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|a Bacterial Proteins
|2 NLM
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|a DNA-Binding Proteins
|2 NLM
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|a DnaA protein, Bacteria
|2 NLM
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|a Recombinant Fusion Proteins
|2 NLM
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|a Acetylglucosaminidase
|2 NLM
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|a EC 3.2.1.52
|2 NLM
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1 |
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|a Froelich, J M
|e verfasserin
|4 aut
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1 |
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|a Phuong, T K
|e verfasserin
|4 aut
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1 |
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|a Forsyth, R A
|e verfasserin
|4 aut
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1 |
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|a Newman, V G
|e verfasserin
|4 aut
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1 |
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|a Zyskind, J W
|e verfasserin
|4 aut
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773 |
0 |
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|i Enthalten in
|t BioTechniques
|d 1993
|g 25(1998), 6 vom: 04. Dez., Seite 1030-5
|w (DE-627)NLM012627046
|x 0736-6205
|7 nnns
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|g volume:25
|g year:1998
|g number:6
|g day:04
|g month:12
|g pages:1030-5
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|d 25
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