The PCR reaction was performed
using Premix EX Taq version 2.0 (Loading dye mix) (Takara)
with the following thermal cycling parameters: 94 ◦C for 3 min, followed
by 35 cycles of 94 ◦C for 30 s, 51 ◦C for 1 min, and 72 ◦C for
2 min, and the final extension was performed at 72 ◦C for 10 min.
The PCR product was cloned into pMD19-T vector (Takara) and
sequenced.