PMC

Intensity of the singlet oxygen emission, obtained by extrapolation of the time-resolved signals back to the laser pulse, from 308 nm flash photolysis of solutions of either 5HT (●) or Rose Bengal (□) in D₂O/H₂O (80%/20% v/v) buffered to pH ∼ 7 with phosphate (40 mmol dm^-3). Inset: Time-resolved luminescence (1270 nm) decays from laser photolysis (308 nm, 10 ns, 5 mJ) of solutions of (a) Rose Bengal, A₃₀₈ = 0.290, and (b) 5HT, A₃₀₈ = 0.266.

Function of A333G mtPheRS in E. coli. Rescue of the growth phenotype of E. coli NP37 transformed with (A) yeast MSF1, msf1-1, or msf1 with a stop codon at A333 and (B) human FARS2, fars2-1, or fars2 with a stop codon at A308. (C) Transediting activity of E. coli wild-type and NP37 FARS2 complemented cell extracts grown at 30 or 42 °C. Reactions were carried out at 42 °C. Data points are an average of three independent experiments with errors bars representing 1 SD.

Coexpression of envelope-genes. HEK293 cells were transfected with pHxBΔCat and pSV-A308T321 (lanes 1, 4, and 8), -A308 (lanes 2, 5, 9) and an equal mixture of the two (lanes 3, 6, and 10). Cell lysates (lanes 1 to 3), virion-depleted supernatants (lanes 4 to 6), and virion pellets (lanes 8 to 10) were prepared 72 h posttransfection and subjected to immunoblotting using as indicated MAb P4D10 specific for HIV-A308 gp120, immune guinea pig serum gp-serum B recognizing both HIV-A308 and HIV-A308T321 gp120, and MAb BC1071 against Gag proteins as detecting reagents. For comparison, a virion-depleted supernatant (lane 7) and a virion pellet (lane 11) derived from a fully infectious HIV-A308 clone propagated in H9 cells were loaded onto the gel. The expression of Envelope protein on the virion was regulated by varying the relative proportion of pSV-A308 to pSV-T321 in the transfection mixture.

Schematic diagrams of transgene constructs used in tobacco and/or tomato transformation. (Right) The predicted structure of RNA transcripts from these transgenes. RPVSat is a full-length (322-nt) sequence of the cereal yellow dwarf virus RPV satellite. The PSTVd sequence in GUS:PSTVd[+] and GUS:PSTVd[-] contains several minor sequence mutations (arising from cloning) compared with the 359-nt wild-type PSTVd-RG1 sequence (deletions of G₁₆₈ and C₃₁₈G₃₁₉ and single-nucleotide substitutions of C₁₈₅→T and C₂₃₁→G), but these mutations do not cause significant alteration to the predicted rod-like structure of the viroid RNA. The sense PSTVd sequence in hpPSTVd starts at nucleotide 16 and ends at nucleotide 355 of the RG1 strain, with one nucleotide substitution of G₂₈₁→A, and the antisense sequence is the same as that in the GUS:PSTVd fusion constructs. There are two versions (A and B) for the hpY-Sat and d/rY-Sat constructs; the Y-SatA sequence has the size of a wild-type Y-Sat RNA (369 nt) but with two single-nucleotide substitutions of A₂₉₃→G and A₃₀₈→G, and Y-SatB has a 5-nt deletion (nucleotides 192-196), which constitutes part of the yellow symptom domain, plus four single-nucleotide substitutions of G₈₀→A, G₁₂₈→ C, G₂₀₄→T, and A₂₁₄→T. The hpGUS sequence is the same as that described in ref. 18. 35S-P, cauliflower mosaic virus 35S promoter; Ocs-T, 3′ region of Agrobacterium octopine synthase gene; i/r, inverted-repeat sequences.