Figure 8.

Accumulation of Double-Stranded gusA DNA in Tobacco BY-2 Protoplasts following Cotransfection with Constructions Expressing Various Histone cDNAs or Mutations of HTA1.

Tobacco protoplasts were cotransfected with a gusA expression construction and either an empty vector control plasmid or constructions expressing various histone cDNAs or histone mutations. At 0 and 24 h after transfection, samples were treated with DNase I and DNA was extracted. PCR was conducted to determine the amount of gusA DNA within the cell, relative to actin DNA. The relative intensities of the ethidium-stained gel bands were determined by densitometric scanning, using Lab Works 4.1 software. Intensity is in arbitrary units, starting at 400. Error bars indicate se of two or three biological replicates.

(A) Empty vector, empty vector + gusA gene; HTA1, HTA1 + gusA gene; HTB1, HTB1 + gusA gene; HTR6, HTR 6 + gusA gene; HTR11, HTR11 + gusA gene; HFO, HFO3 + gusA gene.

(B) Empty vector, empty vector + gusA gene; H2A-1 full-length, full-length H2A-1 + gusA gene; H2A-1 C terminus, C-terminal region of H2A-1 + gusA gene; H2A-1 N terminus, N-terminal region of H2A-1 + gusA gene; H2A-1 N terminus RK→A, first 39 amino acids of H2A-1 in which all basic amino acids were converted to Ala + gusA gene.