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Fig. 6 | AMB Express

Fig. 6

From: Escherichia coli AraJ boosts utilization of arabinose in metabolically engineered cyanobacterium Synechocystis sp. PCC 6803

Fig. 6

Construction of A-Ut5 strain and comparison of growth and sugar consumption between A-Ut1, A-Ut3 and A-Ut5. a araJ was inserted into neutral site 3 under the control of psbA2 promoter (Prm) and 5ST1T2 terminator (Trm) in the A-Ut1 genome, using upstream (N3UP) and downstream (N3DW) regions for homologous recombination. The spectinomycin resistance cassette (Spr) was used for selection and segregation of the transformant. Numbered letters and arrows represent the primer pairs and the direction of the amplification, respectively. b The presence of araJ was verified with the help of primers C1-C2, which generated a ~1.18 kb band. A-Ut1 strain served as the negative control. c Chromosomal segregation was examined with the help of primers G1-G2. In the absence of uninterrupted neutral site 3, the primers failed to generate a PCR product, whereas the A-Ut1 strain included as the positive control produced a ~1.93 kb band. d Transcription of araJ was demonstrated with the help of primers C1-C2, represented as ‘Test’, where the product of ~1.18 kb was obtained. In the positive control (+), petA-specific primers produced a ~0.99 kb band from cDNA template. In the negative control (-), primers C1-C2 failed to amplify araJ from pure RNA sample. e A-Ut1, A-Ut3 and A-Ut5 were grown under mixotrophy in the presence of 20 mM (3.00 g/L) L-arabinose. Dry biomass and residual sugars were measured in the cultures grown in triplicate. (f) A-Ut1, A-Ut3 and A-Ut5 were grown under mixotrophy in the presence of 10 mM (1.50 g/L) L-arabinose and 10 mM (1.80 g/L) D-glucose. Dry biomass and residual sugars were measured in the cultures grown in triplicate

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