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Figure 4 | Microbial Cell Factories

Figure 4

From: Control of total GFP expression by alterations to the 3′ region nucleotide sequence

Figure 4

Comparison of the fluorescence (A) and RT-PCR (B) data from the MK 6 -GFP-Stop(TAA)-(xxx)-# clones and their abbreviations represented by the inserted trinucleotide (in bold type). To obtain the corresponding clones, we inserted single or repeated trinucleotides just beyond the stop codon in the 3′-UTR, outside of the ORF, in the MK6-GFP-Stop(TAA)-(xxx [= the inserted trinucleotide, indicated in lowercase letters])-# clone, as described in the Methods. Determinations of the total protein fluorescence and semi-quantitative RT-PCR were conducted as in Figure 2. Data from products generated after 30 cycles of RT-PCR from the 1-h induced cultures were compared among the clones. The upper and lower bands and the size marker are indicated as in Figure 2. All cultures, mean values, and error bars are as in Figure 1. Lanes: 1, MK6-GFP-LE-6H-Stop(TGA), undeleted control; 2, MK6-GFP-Stop(TAA)-#, deleted control; 3, MK6-GFP-Stop(TAA)-6×taa-#, 6× taa; 4, MK6-GFP-Stop(TAA)-6×ctc-#, 6× ctc; 5, MK6-GFP-Stop(TAA)-1×gag-#, 1× gag; 6, MK6-GFP-Stop(TAA)-3×gag-#, 3× gag; 7, MK6-GFP-Stop(TAA)-6×gag-#, 6× gag; 8, MK6-GFP-Stop(TAA)-6×cac-#, 6× cac; 9, MK6-GFP-Stop(TAA)-6×gaa-#, 6× gaa; 10, MK6-GFP-Stop(TAA)-6×aaa-#, 6× aaa; 11, MK6-GFP-Stop(TAA)-6×aag-#, 6× aag; 12, MK6-GFP-Stop(TAA)-6×ggg-#, 6× ggg; 13, MK6-GFP-Stop(TAA)-6×ttt-#, 6× ttt; 14, MK6-GFP-Stop(TAA)-6×cca-#, 6× cca; 15, MK6-GFP-Stop(TAA)-6×tga-#, 6× tga; and 16, MK6-GFP-Stop(TAA)-6×aga-#, 6× aga.

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