Pet 29a

The bacterial strains and plasmids used in this study are listed in Table 3. The E. coli strains were grown at 37 °C in LB broth or on LB agar. Other bacterial strains were grown aerobically at 30 °C in LB broth or on LB agar. The following antibiotics were used at the indicated concentrations: ampicillin (Ap), 100 μg/mL; spectinomycin (Sm), 100 μg/mL; gentamicin (Gm), 20 μg/mL; and kanamycin (Km), 50 μg/mL.
All of the enzymes used in the DNA manipulations were obtained from TaKaRa Biotechnology Co. Ltd (Dalian, China). E. coli DH5α and BL21 (DE3) were purchased from TaKaRa and were used for cloning and expression hosts, accordingly. The plasmids pMD19-T (TaKaRa) and pET-29a (+) (TaKaRa) were used as cloning and expression vectors, respectively.

Glucose and other cello-oligosaccharide (G₂-G₅), carboxymethyl-cellulose (CMC), barley β-glucan, laminarin, xylan (from corncob), avicel, chitosan, and filter paper were obtained from McLean Biochemical Technology Co., Ltd. (Shanghai, China), whereas lichenin was purchased from Megazyme (Bray, Ireland). Other chemicals used in this study were provided by Sigma-Aldrich (Shanghai, China).
Myxococcus sp. B6-1 was isolated from soil and cultured in a lysogeny broth (LB). DH5 and BL21 (DE3) strains of E. coli from our laboratory were respectively utilized for gene cloning and expression. Plasmid pET-29a (Takara, Beijing, China) was used for expression.