Journal: Metabolic Engineering Communications
Article Title: Isobutanol production in Synechocystis PCC 6803 using heterologous and endogenous alcohol dehydrogenases
Figure Lengend Snippet: Comparison of isobutanol production in engineered Synechocystis PCC 6803 strains Syn-IB-1, -2, -3, -4, -5, and -6. A) RT-PCR, SDS-PAGE and Strep-tag Western-immunoblot (top to bottom). Every dashed line separated each lane represents the results from a single engineered strain. –RT is a negative RT-PCR control using RNA as template without addition of RT enzyme to control for possible DNA contamination. The positive control is an RT-PCR carried out using the corresponding plasmid as template. Red arrows in the SDS-PAGE indicate the location of kivd , blue arrows indicate the expected location for slr1192 . The Strep-tag Western-immunoblot examines the expression of all the ADHs. The negative control for SDS-PAGE and Western-immunoblot is an extract from strain Syn-pDDH. B) Growth curve during 7 days of cultivation. C) Isobutanol production at day 3, day 5 and day 6 from the different engineered strains. Results are the mean of 4 biological replicates and 3 technical replicates, error bars represent standard deviation. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)
Article Snippet: 2.6 RNA isolation and semi-quantitative reverse transcript PCR (RT-PCR) Total RNA was isolated from cultures (OD750 = 0.5) using RTI Reagent (Sigma-Aldrich) according to the manufacturer's instructions.
Techniques: Periodic Counter-current Chromatography, Reverse Transcription Polymerase Chain Reaction, SDS Page, Strep-tag, Western Blot, Positive Control, Plasmid Preparation, Expressing, Negative Control, Standard Deviation