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Adenine deaminase is encoded by Tad1 and participates in copper accumulation in Trichoderma reeseiKehe Fu, LiLi Fan, Chuangjing Yu,Yingying Li, Shigang Gao, Yaqian Li and Jie Chen

Department of Resource and Environmental Science, School of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200240

Fig. S1.

Corresponding author: Tel: +8621-34206141; Fax: +8621-34206141.

E-mail: jiechen59@sjtu.edu.cn (J. Chen)

Schematic representation of the construction of plasmids. A, plasmid of Tad1 overexpression. B,

plasmid of RNA-mediated Tad1 silencing.

Fig. S2.

Southern blotting analysis of the mutants to confirm the copies of T-DNA insertion. P, plasmid for

positive control. Ov-1 to Ov-3, three random select mutants of overexpression of Tad1. RNAi-1 to

RNAi-3, three random select mutants of RNA silent of Tad1. For southern blotting, the genomes DNA

of mutants were digested with XbaI and 0.9% agarose gel. Fragment of ORF of hph was set as probe.

Hybridization was carried out according to Gene Images CDP-Star Detection Kit (Amersham, UK).

Fig. S3.

mRNA expression profiles of Tad1 in WT and mutants. △promTad1, a mutant deleted 500 bp DNA

sequence before -707 bp upstream of T-DNA insertion in AT01. ATCK, a mutant contained an

integrated T-DNA cassette which inserted in the same site as AT01.

Fig. S4.

Graphical representation of how Tad1 regulated the copper homeostasis in T.reesei according to the

copper homeostasis in S.cerevisiae. DMT, hypothetical transmembrane divalent metal ions transporter.

Ace1, copper depend transcription factor. MTs, metallothioneins. Atx1, Cccs1 and Cox17 were copper

chaperones.