Biochemical construction and selection of hybrid plasmids containing specific segments of the Escherichia coli genome.
نویسندگان
چکیده
منابع مشابه
Biochemical construction of specific chimeric plasmids from ColE1 DNA and unfractionated Escherichia coli DNA.
A series of chimeric plasmids was constructed using colicinigenic factor E1 (ColE1) DNA as the replicon and DNA fragments carrying the galactose or tryptophan operons from E. coli. Restriction endonuclease EcoRI digests of ColE1 DNA and various DNAs containing the trp or gal operons were joined by T4 polynucleotide ligase [polynucleotide synthetase (ATP), poly(deoxyribonucleotide):poly(deoxyrib...
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ESCHERICHIA COLI HEAT-LABILE TOXIN B SUBUNIT: CONSTRUCTION AND EVALUATION OF PLASMIDS PROVIDING CONTROLLED HIGH LEVEL PRODUCTION OF THE PROTEIN
With the plasmid DNA from a clinical isolate of enterotoxigenic Escherichia coli (ETEC) H 10407 as template, PCR-mediated cloning of the sequence encoding the heat-labile toxin B subunit (L T -B) has been carried out. Then this sequence was recloned into the pTrc 99A and pET23a expression vectors to give the pJasmids pTRCLTB and pETLTB, respectively. After induction, the former plasmid provides...
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In order to study the periplasmic expression of human growth hormone (hGH) in Escherichia coli, the related cDNA was inserted in two expression plasmids carrying pelB signal peptide, one with lac bacterial promoter and the other with a bacteriophage T7-based promoter. The recombinant plasmids were moved to TG1 and BL21 strains of E. coli, respectively. To induce the expression systems, IPTG and...
متن کاملIdentification of the Escherichia coli tonB gene product in minicells containing tonB hybrid plasmids.
Sotlillm clodtq~l sul~~t~e/polyacrylarnide gel :tnwlysis ot’ protrins t~codcd by ii sc>rics ot’ to?/ R + plasmids in minicells has icknt~ified the torLB gene product ws a prokin wit)h km apparent, molecular weight of’ 36,000. A parallel analysis of s(‘vcn tort N mnt,ations which have been genetically crosst~i onto a tonB+ plasmid support,s this itkntification; t)he 36,000 M, protein is tlbsent ...
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ژورنال
عنوان ژورنال: Proceedings of the National Academy of Sciences
سال: 1975
ISSN: 0027-8424,1091-6490
DOI: 10.1073/pnas.72.11.4361