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PMID: 6212753 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Amplification of maize ribulose bisphosphate carboxylase large subunit synthesis in E. coli by transcriptional fusion with the lambda N operon.

Molecular & general genetics : MGG ·Vol. 185 ·No. 3 ·1982-00-00 ·Pages 424-9

Gatenby AA, Castleton JA

Abstract

The maize chloroplast gene coding for the large subunit of ribulose bisphosphate carboxylase (3-phospho-D-glycerate carboxy-lyase (dimerizing), EC 4.1.1.39) has been placed under the transcriptional control of the bacteriophage lambda promoter PL, by fusion with the lambda N operon located on a multicopy plasmid. Transcription from PL was repressed at 32 degrees C by the presence in the E. coli chromosome of a cIts gene that specifies a temperature-sensitive repressor. After inactivation of the repressor at 45 degrees C unmoderated transcription of the chloroplast gene occurred from the PL promoter. Translation was probably initiated from a chloroplast Shine-Dalgarno sequence located five nucleotides from the N-terminal methionine initiation codon to yield a polypeptide the same size as that synthesised in maize. This direct translation results in a level of expression of the chloroplast gene corresponding to approximately 2% of the total E. coli cell protein as ribulose bisphosphate carboxylase large subunits. Transcriptional fusions with the lambda N operon should provide a generally applicable, simple method for the amplification and regulation of chloroplast gene expression in E. coli.

MeSH Terms
Bacteriophage lambda/genetics Carboxy-Lyases/genetics Escherichia coli/genetics Gene Amplification Kinetics Operon Plants/enzymology,genetics Plasmids Ribulose-Bisphosphate Carboxylase/genetics Transcription, Genetic Zea mays/enzymology,genetics
Chemicals
Carboxy-Lyases Ribulose-Bisphosphate Carboxylase
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Gatenby A A
Castleton J A
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30 references, click to expand
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Article Info
Journal
Molecular & general genetics : MGG
Abbr.
Mol Gen Genet
ISSN
0026-8925
DOI
10.1007/BF00334134
Published
1982-00-00
Pages
424-9
Language
English
Region
Germany
NLM ID
0125036
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00334134
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