Extended Data Fig. 2: Nucleic acid melting activity assay of wheat TaCSP-H and TaCSP-V.
From: Horizontally acquired CSP genes contribute to wheat adaptation and improvement
a. Nucleic acid unfolding activity assay of TaCSP-H1 from all three sub-genomes of wheat in E. coli BX04 strain. Diluted cultures of BX04 cells expressing bacterial CspA, PINIII (empty vector) or TaCSP-H1 were spotted onto LB-carbenicillin plates and incubated at 15 °C or 37 °C. CspA and PINIII were used as the positive and negative controls, respectively. The mRNA levels indicate the expression of genes used in the assay. b. Transcription anti-termination assay of TaCSP-H2 and TaCSP-H3 in E. coli RL211 strain. Diluted cultures of RL211 cells expressing TaCSP-H2 and TaCSP-H3 were spotted onto LB plates with ( + Cm) or without (-Cm) chloramphenicol. CspA and PINIII were used as the positive and negative controls, respectively. The mRNA levels indicate the expression of genes used in the assay. c-e. Transcriptional anti-termination assay of the TaCSP-V in E. coli RL211 strain. Diluted cultures of RL211 cells expressing TaCSP-V1 (TraesCS1D03G0632700), TaCSP-V2 (TraesCS1D03G0633000), TaCSP-V3 (TraesCS6D03G0140500) and their CSD domain truncations (TaCSP-V-CSD) were spotted onto LB plates with ( + Cm) or without (-Cm) chloramphenicol. CspA and PINIII were used as the positive and negative controls, respectively. The mRNA levels indicate the expression of genes or gene truncations used in the assay.
