Extended Data Fig. 5: Allotopic expression of Nad9.
From: Targeted knockout of a conserved plant mitochondrial gene by genome editing
(a) Structure of plant transformation vector pJF1271 used for allotopic complementation. LB, T-DNA left border; PAtUBQ10, promoter from the Arabidopsis thaliana UBIQUITIN10 gene; StFDH, Solanum tuberosum FORMATE DEHYDROGENASE; TAtUBQ10, terminator from the Arabidopsis UBIQUITIN10 gene; Pnos, promoter from the Agrobacterium tumefaciens nopaline synthase-encoding gene; hpt (hygR), gene encoding hygromycin phosphotransferase conferring resistance to hygromycin; Tnos, terminator from the Agrobacterium gene encoding nopaline synthase; RB, T-DNA right border; ori (pSa) origin of replication in A. tumefaciens; specR, spectinomycin resistance; ori (pUC) origin of replication in E. coli. (b) Analysis of the expression level of the nuclear nad9 copy in the complemented lines by semiquantitative RT-PCR. β-TUBULIN expression served as internal control. The numbers at the bottom of the lanes indicate the relative intensity of the nad9 signal (given as percentage of the total signal). M, DNA size marker; +RT, reverse-transcribed RNA; -RT, control reactions without addition of reverse transcriptase; H2O, buffer control; cyan arrowhead, β-TUBULIN RT-PCR product; magenta arrowhead, nad9 RT-PCR product. In the strict sense, the experiments presented here were not repeated independently.
