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Fig. 6 | Microbial Cell Factories

Fig. 6

From: The expression and construction of engineering Escherichia coli producing humanized AluY RNAs

Fig. 6

The diagrammatic sketch of plasmid construction and enzyme digestion. a Schematic diagram of pEGFP-C1 derived vectors. The tandem antisense AluY sequences (AluY × 1as, AluY × 2as AluY × 4as AluY × 8as AluY × 14as) were inserted the multiple cloning site (MCS) of pEGFP-C1 vector. The order of restriction enzyme cutting sites in pEGFP-C1 vector was 5′-HindIII site-antisense Alus-NheI site-3′, the arrows show the orientation of transcription. b Schematic diagram of pET derived vectors. Tandem antisense AluY sequences (relative to pEGFP-C1) digested from pEGFP-C1 derived vectors (C1-AluY × 1as, C1-AluY × 2as, C1-AluY × 4as C1-AluY × 8as, C1-AluY × 14as) were inserted into pET. NheI site in pET vector is located downstream of T7 promoter so that antisense AluYs in pEGFP-C1 inserted into pET vector transcribed sense AluY RNAs. The order of restriction enzyme cutting sites in pET vector was 5′-T7 promoter-NheI site-Alus-HindIII site-3′. c Agarose gel electrophoresis images of plasmids being digested with HindIII/NheI (ethidium bromide staining). Lane 1: pET-AluY × 1, lane 2: pET-AluY × 2, lane 3: pET-AluY × 4, lane 4: pET-AluY × 8, lane 5: pET-AluY × 14, lane 6: marker

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