scholarly journals Transient Conversion to RecA+ Phenotype to Permit P1 Transduction in any Escherichia coli recA- Strains

BioTechniques ◽  
1999 ◽  
Vol 27 (5) ◽  
pp. 911-914 ◽  
Author(s):  
Marian Sektas ◽  
Magdalena Gregorowicz ◽  
Waclaw Szybalski
Keyword(s):  
Escherichia Coli ◽  
P1 Transduction

scholarly journals Triple knockout of frdC gltA and pta genes enhanced pHA production in Escherichia coli

2018 ◽  
pp. 11-18
Author(s):  
Nurhajirah Mohamed Biran ◽  
Mohd Zulkhairi Mohd Yusoff ◽  
Toshinari Maeda ◽  
Mohd Rafein Zakaria ◽  
Lian-Ngit Yee ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Pha Synthase ◽  
Control Strain ◽  
E Coli ◽  
Pha Production ◽  
Coa Reductase ◽  
Genome Information ◽  
Acetoacetyl Coa Reductase ◽  
P1 Transduction ◽  
Engineered Strain

Polyhydroxyalkanoate (PHA) is a linear polyester produced through the fermentation of sugar or lipid. Biosynthesis of PHA comprises three enzymes known as acetyl-CoA acetyltransferase (phaA), acetoacetyl-CoA reductase (phaB) and PHA synthase (phaC). Comamonas sp. is one of the strains commonly used for PHA production. In order to develop higher PHA production from bacterial respond strategy, PHA biosynthesis operon of Comamonas sp. EB172 was introduced into Escherichia coli BW25113 through a pGEM-T vector. E. coli was chosen due to the complete genome information available and the absence of depolymerisation gene, phaZ. In this study, the deletion of several single genes, which are frdC, gltA, and pta, was found to be associated with PHA metabolism activity in E. coli BW25113. P1 transduction was performed to construct multiple genes knockout. The engineered strain, E. coli BW25113 frdCgltApta::kan/pGEM’-phaCABCo, yielded the highest PHA production at 64 wt.% with 1.4 fold higher than that of control strain of E. coli BW25113/pGEM’-phaCABCo. This strain is potential for industrial application for higher PHA production from E. coli.

scholarly journals GENETIC ANALYSIS OF AN ESCHERICHIA COLI MUTANT WITH A LESION IN STABLE RNA TURNOVER

Genetics ◽  
1974 ◽  
Vol 76 (2) ◽  
pp. 185-194
Author(s):  
Yoshinari Ohnishi
Keyword(s):  
Escherichia Coli ◽  
Genetic Analysis ◽  
Gene Order ◽  
Rna Turnover ◽  
E Coli ◽  
Coli Mutant ◽  
Its Rrna ◽  
Derivatives Of ◽  
P1 Transduction

ABSTRACT A mutant that rapidly degrades more than 80% of its rRNA and tRNA under defined conditions was genetically analyzed. Two genes, srnA and srnB, are separately located, and the mutated alleles of both are required for degradation of stable RNA in cultures treated with rifampicin at 42°. srnA is closely linked to tsx by matings and transduction tests; by P1 transduction, the gene order is lac (9 min) proC (9.55 min) tsx (9.8 min) srnA (about 10 min) purE (12 min) rnsA (14.4 min). srnB is not yet completely mapped, but is outside the lac-rnsA region, probably in the region between 75 and 90 min.—The product of the rnsA gene, RNase I, is a potent endonuclease of E. coli, and the only one known that can attack ribosomes and tRNA. However, not only are the srn lesions genetically separate from rnsA, but also, derivatives of an srn strain were prepared lacking RNase I, and they retain the Srn- phenotype. Thus, no correlation of rapid RNA turnover and RNase I activity has been found.

scholarly journals Further Studies with Lipoamide Dehydrogenase Mutants of Escherichia coli k12

Microbiology ◽  
2000 ◽  
Vol 81 (1) ◽  
pp. 237-245 ◽  
Author(s):  
J. R. Guest ◽  
I. T. Creaghan
Keyword(s):  
Escherichia Coli ◽  
Pyruvate Dehydrogenase ◽  
Recombination Frequency ◽  
Regulatory Element ◽  
Multienzyme Complex ◽  
Wild Type ◽  
Mutant Site ◽  
Lipoamide Dehydrogenase ◽  
Distal Gene ◽  
P1 Transduction

The immunological properties of ten lipoamide dehydrogenase mutants of Escherichia coli were investigated with antiserum raised against purified lipoamide dehydrogenase. Seven mutants were CRM+ (cross-reacting material present) as they contained lipoamide dehydrogenase proteins exhibiting either complete or partial immunological identity with the wild-type protein. This indicates that at least seven of the mutations affect the lipoamide dehydrogenase structural gene (lpd). The remaining three mutants (CRM-) contained no detectable cross-reacting protein. None of the lpd mutations were sensitive to any of three different amber-suppressors. Genetic analysis by P1-transduction showed that all the lpd mutant sites were clustered very near the distal gene (aceF) of the ace region which specifies the dehydrogenase (aceE) and transacetylase (aceF) components of the pyruvate dehydrogenase multienzyme complex. Calculations based on the recombination frequency between an aceF mutant and the nearest lpd mutant site support the conclusion that apart from the possible presence of a regulatory element, the aceF and lpd genes are contiguous.

scholarly journals Positive and negative selection using the tetA-sacB cassette: recombineering and P1 transduction in Escherichia coli

2013 ◽  
Vol 41 (22) ◽  
pp. e204-e204 ◽  
Author(s):  
Xin-tian Li ◽  
Lynn C. Thomason ◽  
James A. Sawitzke ◽  
Nina Costantino ◽  
Donald L. Court
Keyword(s):  
Escherichia Coli ◽  
Negative Selection ◽  
P1 Transduction

Construction of an Escherichia coli strain which excretes abnormally large amounts of adenosine 3′,5′-cyclic monophosphate

1978 ◽  
Vol 24 (11) ◽  
pp. 1423-1425 ◽  
Author(s):  
Ann D. E. Fraser ◽  
Hiroshi Yamazaki
Keyword(s):  
Escherichia Coli ◽  
Minimal Medium ◽  
Coli Strain ◽  
Coli Cell ◽  
Wild Type ◽  
Camp Phosphodiesterase ◽  
E Coli ◽  
Cyclic Monophosphate ◽  
Glucose Minimal Medium ◽  
P1 Transduction

It has previously been shown that an Escherichia coli CRP− strain 5333 accumulates abnormally large amounts of adenosine 3′,5′-cyclic monophosphate (cAMP). Using P1 transduction, the CRP− character was transferred to E. coli Crookes strain which is deficient for cAMP phosphodiesterase (CPD−). The resulting strain HY22 (CRP−, CPD−) accumulates greater amounts of cAMP both intracellularly and extracellularly than does 5333. In glucose minimal medium, an HY22 cell accumulates 100 times more cAMP intracellularly and excretes cAMP 150 times faster than does a wild-type E. coli cell.

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