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. 2020 Sep 10;11:552160. doi: 10.3389/fpls.2020.552160

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Copyright © 2020 Okada, Gregg, Allen, Menon, Hussain, Gillespie, Johnston, Byrne, Colgrave and Wood

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Design of the pFAγ51 MTP and assessment of its cleavage when fused to different Nif proteins expressed in plants. (A) Schematic of the truncation of the N-terminal amino acid sequence of the A. thaliana F1 ATPase γ subunit from 77 to 51 residues. The 26 amino acid residues in the orange section of pFAγ were removed. MTP, Nif, HA epitope tag are not to scale. MTP, mitochondrial targeting peptide; MPP, mitochondrial processing peptidase. (B) Western blot analysis (α-HA) of of the total protein extracts of individual pFAγ51::Nif::HA, pFAγ51::HA::NifK, 6×His::Nif::HA and 6×His::HA::NifK proteins transiently expressed in N. benthamiana leaf. Black dots point to the size of the correctly processed pFAγ51::Nif::HA protein. C, cytosolic expression; M, mitochondrial targeted. Panels of individual Nif proteins shown in B were extracted from full blot images presented in Supplementary Figure 2 .