FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2950470251> ?p ?o ?g. }

• W2950470251 abstract "ABSTRACT Collections of characterized promoters of different strengths are key resources for synthetic biology, but are not well established for many important organisms, including industrially-relevant Clostridium spp. When generating promoters, reporter constructs are used to measure expression, but classical fluorescent reporter proteins are oxygen-dependent and hence inactive in anaerobic bacteria like Clostridium . We directly compared oxygen-independent reporters of different types in Clostridium acetobutylicum and found that glucuronidase (GusA) from E. coli performed best. Using GusA, a library of synthetic promoters was first generated by a typical approach entailing complete randomization of a constitutive thiolase gene promoter (P thl ) except for the consensus -35 and -10 elements. In each synthetic promoter, the chance of each degenerate position matching P thl was 25%. Surprisingly, none of the synthetic promoters from this library were functional in C. acetobutylicum , even though they functioned as expected in E. coli . Next, instead of complete randomization, we specified lower promoter mutation rates using oligonucleotide primers synthesized using custom mixtures of nucleotides. Using these primers, two promoter libraries were constructed in which the chance of each degenerate position matching P thl was 79% or 58%, instead of 25% as before. Synthetic promoters from these ‘stringent’ libraries functioned well in C. acetobutylicum , covering a wide range of strengths. The promoters functioned similarly in the distantly-related species Clostridium sporogenes , and allowed predictable metabolic engineering of C. acetobutylicum for acetoin production. Besides generating the desired promoters and demonstrating their useful properties, this work indicates an unexpected ‘stringency’ of promoter sequences in Clostridium , not reported previously. GRAPHICAL ABSTRACT" @default.
• W2950470251 created "2019-06-27" @default.
• W2950470251 creator A5080805114 @default.
• W2950470251 creator A5084171210 @default.
• W2950470251 date "2017-11-09" @default.
• W2950470251 modified "2023-09-23" @default.
• W2950470251 title "Stringency of synthetic promoter sequences inClostridiumrevealed and circumvented by tuning promoter library mutation rates" @default.
• W2950470251 cites W1633114099 @default.
• W2950470251 cites W1834065866 @default.
• W2950470251 cites W1963562346 @default.
• W2950470251 cites W1964772492 @default.
• W2950470251 cites W1975765580 @default.
• W2950470251 cites W1978693132 @default.
• W2950470251 cites W1980623154 @default.
• W2950470251 cites W1991383594 @default.
• W2950470251 cites W1996829602 @default.
• W2950470251 cites W1998585003 @default.
• W2950470251 cites W2000623234 @default.
• W2950470251 cites W2002127274 @default.
• W2950470251 cites W2008016941 @default.
• W2950470251 cites W2016036293 @default.
• W2950470251 cites W2018627617 @default.
• W2950470251 cites W2026062081 @default.
• W2950470251 cites W2027908623 @default.
• W2950470251 cites W2033345778 @default.
• W2950470251 cites W2036431748 @default.
• W2950470251 cites W2047821263 @default.
• W2950470251 cites W2057141078 @default.
• W2950470251 cites W2076243547 @default.
• W2950470251 cites W2076701025 @default.
• W2950470251 cites W2085970397 @default.
• W2950470251 cites W2088710570 @default.
• W2950470251 cites W2090591880 @default.
• W2950470251 cites W2094522474 @default.
• W2950470251 cites W2098103683 @default.
• W2950470251 cites W2099047627 @default.
• W2950470251 cites W2101024694 @default.
• W2950470251 cites W2104059792 @default.
• W2950470251 cites W2110146873 @default.
• W2950470251 cites W2111967267 @default.
• W2950470251 cites W2119225291 @default.
• W2950470251 cites W2125905611 @default.
• W2950470251 cites W2134783465 @default.
• W2950470251 cites W2138638690 @default.
• W2950470251 cites W2140373206 @default.
• W2950470251 cites W2146205446 @default.
• W2950470251 cites W2146372898 @default.
• W2950470251 cites W2151530177 @default.
• W2950470251 cites W2152007804 @default.
• W2950470251 cites W2153799311 @default.
• W2950470251 cites W2154317105 @default.
• W2950470251 cites W2157897258 @default.
• W2950470251 cites W2158849520 @default.
• W2950470251 cites W2165924947 @default.
• W2950470251 cites W2210361504 @default.
• W2950470251 cites W2223848185 @default.
• W2950470251 cites W2304664643 @default.
• W2950470251 cites W2325347923 @default.
• W2950470251 cites W2382044701 @default.
• W2950470251 cites W2399213131 @default.
• W2950470251 cites W2412647772 @default.
• W2950470251 cites W2418004535 @default.
• W2950470251 cites W2508072828 @default.
• W2950470251 cites W2509283163 @default.
• W2950470251 cites W2522630305 @default.
• W2950470251 cites W2538295181 @default.
• W2950470251 cites W2622258133 @default.
• W2950470251 cites W2625319875 @default.
• W2950470251 cites W297084931 @default.
• W2950470251 cites W4239222085 @default.
• W2950470251 doi "https://doi.org/10.1101/216853" @default.
• W2950470251 hasPublicationYear "2017" @default.
• W2950470251 type Work @default.
• W2950470251 sameAs 2950470251 @default.
• W2950470251 citedByCount "0" @default.
• W2950470251 crossrefType "posted-content" @default.
• W2950470251 hasAuthorship W2950470251A5080805114 @default.
• W2950470251 hasAuthorship W2950470251A5084171210 @default.
• W2950470251 hasBestOaLocation W29504702511 @default.
• W2950470251 hasConcept C101762097 @default.
• W2950470251 hasConcept C104317684 @default.
• W2950470251 hasConcept C150194340 @default.
• W2950470251 hasConcept C161733203 @default.
• W2950470251 hasConcept C191908910 @default.
• W2950470251 hasConcept C2776928978 @default.
• W2950470251 hasConcept C2777713963 @default.
• W2950470251 hasConcept C2780161600 @default.
• W2950470251 hasConcept C2781092032 @default.
• W2950470251 hasConcept C523546767 @default.
• W2950470251 hasConcept C54355233 @default.
• W2950470251 hasConcept C55493867 @default.
• W2950470251 hasConcept C70721500 @default.
• W2950470251 hasConcept C86803240 @default.
• W2950470251 hasConceptScore W2950470251C101762097 @default.
• W2950470251 hasConceptScore W2950470251C104317684 @default.
• W2950470251 hasConceptScore W2950470251C150194340 @default.
• W2950470251 hasConceptScore W2950470251C161733203 @default.
• W2950470251 hasConceptScore W2950470251C191908910 @default.
• W2950470251 hasConceptScore W2950470251C2776928978 @default.
• W2950470251 hasConceptScore W2950470251C2777713963 @default.

• next