Artificial optimization of bamboo transposase and host factors effects on transposition in yeast.

() are promising tools for gene cloning, gene expression, and gene tagging. We have characterized two transposons from moso bamboo, and . , is smaller in size and has higher natural activities, thus making it a more potential genomic tool compared to . Using a two-component system consisting of a transposase expression cassette and a non-autonomous transposon cotransformed in yeast, we investigated the transposition activity of and created hyperactive transposases. Five out of 19 amino acid mutations in outperformed the wild-type in terms of catalytic activities, especially with the S347R mutant having 6.7-fold higher transposition activity. Moreover, 36 yeast mutants with single-gene deletion were chosen to screen the effects of the host factors on transposition. Compared to the control strain (), the mobility of was greatly increased in 9 mutants and dramatically decreased in 7 mutants. The transposition ability in the mutant was 15-fold higher than in the control, while it was lowered to 1/66 in the mutant. Transcriptomic analysis exhibited that defection led to the significantly impaired , expression and dramatically boosted expression, whereas defection resulted in significantly suppressed expression of , and . Protein methylation, chromatin and RNA transcription may affect the transposition efficiency in yeast. Overall, the findings provided evidence for transposition regulation and offered an alternative genomic tool for moso bamboo and other plants.