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. 2017 Apr 18;14(7):926–937. doi: 10.1080/15476286.2017.1311455

Figure 3.

Figure 3.

In- cell solubility of Tat protein in E.coli by TAR RNA. The effect of TAR RNA enhances Tat protein co-expression on the solubility in vivo. (A) The expression of wild type Tat was controlled by T7 promoter and induced by IPTG, whereas the expression of wt TAR RNA was under the control of arabinose promoter and induced by L-arabinose. The band intensity of each Tat protein fraction on the SDS-PAGE gel was estimated by densitometric scanning. T, S, and P represent total lysate, soluble fraction, and pellet fraction, respectively. (B) Increase of Tat-EGFP solubility by TAR co-expression as analyzed by Coomassie staining and western blot using the anti-His antibody. (C) SDS-PAGE gel analysis of Tat mutants. The relative solubility of Tat mutants based on the Coomassie stained band intensities is shown in bars: with (white) and without (black) TAR RNA coexpression. The band intensity of each Tat protein fraction was measured by a densitometer. The mutants M1, M2, and M3 Tat carry amino acid substitutions, R52A, R53A, and both R52A and R53A, respectively. T, S, and P represent total lysate, soluble fraction, and pellet fraction.