FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W4210347382> ?p ?o ?g. }

• W4210347382 abstract "The widespread usage of protein expression systems in Escherichia coli (E. coli) is a workhorse of molecular biology research that has practical applications in biotechnology industry, including the production of pharmaceutical drugs. Various factors can strongly affect the successful construction and stable maintenance of clones and the resulting biosynthesis levels. These include an appropriate selection of recombinant hosts, expression systems, regulation of promoters, the repression level at an uninduced state, growth temperature, codon usage, codon context, mRNA secondary structure, translation kinetics, the presence/absence of chaperons and others. However, optimization of the growth medium's composition is often overlooked. We systematically evaluate this factor, which can have a dramatic effect on the expression of recombinant proteins, especially those which are toxic to a recombinant host.Commonly used animal tissue- and plant-based media were evaluated using a series of clones in pET vector, containing expressed Open Reading Frames (ORFs) with a wide spectrum of toxicity to the recombinant E. coli: (i) gfpuv (nontoxic); (ii) tp84_28-which codes for thermophilic endolysin (moderately toxic); and (iii) tthHB27IRM-which codes for thermophilic restriction endonuclease-methyltransferase (REase-MTase)-RM.TthHB27I (very toxic). The use of plant-derived peptones (soy peptone and malt extract) in a culture medium causes the T7-lac expression system to leak. We show that the presence of raffinose and stachyose (galactoside derivatives) in those peptones causes premature and uncontrolled induction of gene expression, which affects the course of the culture, the stability of clones and biosynthesis levels.The use of plant-derived peptones in a culture medium when using T7-lac hybrid promoter expression systems, such as Tabor-Studier, can lead to uncontrolled production of a recombinant protein. These conclusions also extend to other, lac operator-controlled promoters. In the case of proteins which are toxic to a recombinant host, this can result in mutations or deletions in the expression vector and/or cloned gene, the death of the host or highly decreased expression levels. This phenomenon is caused by the content of certain saccharides in plant peptones, some of which (galactosides) may act as T7-lac promoter inducer by interacting with a Lac repressor. Thus, when attempting to overexpress toxic proteins, it is recommended to either not use plant-derived media or to use them with caution and perform a pilot-scale evaluation of the derepression effect on a case-by-case basis." @default.
• W4210347382 created "2022-02-08" @default.
• W4210347382 creator A5004478824 @default.
• W4210347382 creator A5032601667 @default.
• W4210347382 creator A5040955963 @default.
• W4210347382 creator A5055848460 @default.
• W4210347382 creator A5068456789 @default.
• W4210347382 creator A5073773766 @default.
• W4210347382 date "2022-01-28" @default.
• W4210347382 modified "2023-10-14" @default.
• W4210347382 title "T7-lac promoter vectors spontaneous derepression caused by plant-derived growth media may lead to serious expression problems: a systematic evaluation" @default.
• W4210347382 cites W1543403718 @default.
• W4210347382 cites W1668599603 @default.
• W4210347382 cites W1928833223 @default.
• W4210347382 cites W1975993749 @default.
• W4210347382 cites W1976886327 @default.
• W4210347382 cites W1990619608 @default.
• W4210347382 cites W1992110267 @default.
• W4210347382 cites W2002514085 @default.
• W4210347382 cites W2005411001 @default.
• W4210347382 cites W2012633290 @default.
• W4210347382 cites W2032619610 @default.
• W4210347382 cites W2045732079 @default.
• W4210347382 cites W2051064236 @default.
• W4210347382 cites W2062579188 @default.
• W4210347382 cites W2063027152 @default.
• W4210347382 cites W2064492100 @default.
• W4210347382 cites W2065496171 @default.
• W4210347382 cites W2068518785 @default.
• W4210347382 cites W2076625692 @default.
• W4210347382 cites W2082092933 @default.
• W4210347382 cites W2088144279 @default.
• W4210347382 cites W2100837269 @default.
• W4210347382 cites W2127068865 @default.
• W4210347382 cites W2146581807 @default.
• W4210347382 cites W2154192967 @default.
• W4210347382 cites W2156185271 @default.
• W4210347382 cites W2161420671 @default.
• W4210347382 cites W2162095292 @default.
• W4210347382 cites W2162362206 @default.
• W4210347382 cites W2766989338 @default.
• W4210347382 cites W2796190521 @default.
• W4210347382 cites W2980830843 @default.
• W4210347382 cites W3022637875 @default.
• W4210347382 cites W4244681315 @default.
• W4210347382 cites W4249520856 @default.
• W4210347382 doi "https://doi.org/10.1186/s12934-022-01740-5" @default.
• W4210347382 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/35090462" @default.
• W4210347382 hasPublicationYear "2022" @default.
• W4210347382 type Work @default.
• W4210347382 citedByCount "4" @default.
• W4210347382 countsByYear W42103473822022 @default.
• W4210347382 countsByYear W42103473822023 @default.
• W4210347382 crossrefType "journal-article" @default.
• W4210347382 hasAuthorship W4210347382A5004478824 @default.
• W4210347382 hasAuthorship W4210347382A5032601667 @default.
• W4210347382 hasAuthorship W4210347382A5040955963 @default.
• W4210347382 hasAuthorship W4210347382A5055848460 @default.
• W4210347382 hasAuthorship W4210347382A5068456789 @default.
• W4210347382 hasAuthorship W4210347382A5073773766 @default.
• W4210347382 hasBestOaLocation W42103473821 @default.
• W4210347382 hasConcept C104317684 @default.
• W4210347382 hasConcept C141231307 @default.
• W4210347382 hasConcept C150194340 @default.
• W4210347382 hasConcept C151730666 @default.
• W4210347382 hasConcept C186310378 @default.
• W4210347382 hasConcept C2778597767 @default.
• W4210347382 hasConcept C2779343474 @default.
• W4210347382 hasConcept C2779998883 @default.
• W4210347382 hasConcept C33161422 @default.
• W4210347382 hasConcept C40767141 @default.
• W4210347382 hasConcept C54355233 @default.
• W4210347382 hasConcept C547475151 @default.
• W4210347382 hasConcept C55493867 @default.
• W4210347382 hasConcept C73766848 @default.
• W4210347382 hasConcept C86803240 @default.
• W4210347382 hasConcept C87253356 @default.
• W4210347382 hasConceptScore W4210347382C104317684 @default.
• W4210347382 hasConceptScore W4210347382C141231307 @default.
• W4210347382 hasConceptScore W4210347382C150194340 @default.
• W4210347382 hasConceptScore W4210347382C151730666 @default.
• W4210347382 hasConceptScore W4210347382C186310378 @default.
• W4210347382 hasConceptScore W4210347382C2778597767 @default.
• W4210347382 hasConceptScore W4210347382C2779343474 @default.
• W4210347382 hasConceptScore W4210347382C2779998883 @default.
• W4210347382 hasConceptScore W4210347382C33161422 @default.
• W4210347382 hasConceptScore W4210347382C40767141 @default.
• W4210347382 hasConceptScore W4210347382C54355233 @default.
• W4210347382 hasConceptScore W4210347382C547475151 @default.
• W4210347382 hasConceptScore W4210347382C55493867 @default.
• W4210347382 hasConceptScore W4210347382C73766848 @default.
• W4210347382 hasConceptScore W4210347382C86803240 @default.
• W4210347382 hasConceptScore W4210347382C87253356 @default.
• W4210347382 hasFunder F4320335039 @default.
• W4210347382 hasIssue "1" @default.
• W4210347382 hasLocation W42103473821 @default.
• W4210347382 hasLocation W42103473822 @default.
• W4210347382 hasLocation W42103473823 @default.
• W4210347382 hasLocation W42103473824 @default.
• W4210347382 hasOpenAccess W4210347382 @default.

• next