CRIMoClo plasmids for modular assembly and orthogonal chromosomal integration of synthetic circuits in Escherichia coli
نویسندگان
چکیده
منابع مشابه
Chromosomal transformation of Escherichia coli recD strains with linearized plasmids.
Wild-type Escherichia coli are resistant to genetic transformation by purified linear DNA, probably in part because of exonuclease activity. We demonstrate that E. coli containing a recD mutation could be easily transformed by linearized plasmids containing a selectable marker. The marker was transferred to the chromosome by homologous recombination, whereas plasmid markers not in the region of...
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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 کاملNucleotide sequence of the Escherichia coli recJ chromosomal region and construction of recJ-overexpression plasmids.
The nucleotide sequence of the recJ gene of Escherichia coli K-12 and two upstream coding regions was determined. Three regions were identified within these two upstream genes that exhibited weak to moderate promoter activity in fusions to the galK gene and are candidates for the recJ promoter. recJ appeared to be poorly translated: the recJ nucleotide sequence revealed a suboptimal initiation ...
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Background: The chromosomal integration of biological parts in the host genome enables the engineering of plasmid-free stable strains with single-copy insertions of the desired gene networks. Although different integrative vectors were proposed, no standard pre-assembled genetic tool is available to carry out this task. Synthetic biology concepts can contribute to the development of standardize...
متن کاملA standard vector for the chromosomal integration and characterization of BioBrick™ parts in Escherichia coli
BACKGROUND The chromosomal integration of biological parts in the host genome enables the engineering of plasmid-free stable strains with single-copy insertions of the desired gene networks. Although different integrative vectors were proposed, no standard pre-assembled genetic tool is available to carry out this task. Synthetic biology concepts can contribute to the development of standardized...
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ژورنال
عنوان ژورنال: Journal of Biological Engineering
سال: 2019
ISSN: 1754-1611
DOI: 10.1186/s13036-019-0218-8