... werepurchased from Sigma.DNA manipulation and sequence analysisPlasmid DNA preparation, purification of DNA from agarose gel, and restriction enzyme analysis were performedby the standard methods ... 11718–11728.27. Watanabe,S.,Muramatsu,T.,Ao,H.,Hirayama,Y.,Takahashi,K., Tanokura, M. & Kuchino, Y. (1999) Molecular cloning of theLonproteasegenefromThermus thermophilus HB8 and char-acterization of ... kDa), and carbonic anhydrase (29 kDa)] is shownas the standard curve. The molecular mass of native Bt-Lon (s)wasestimated from the standard curve based on the elution volume of native Bt-Lon and...