pANC209 (xylA)

Protein Expression Prokaryotic cells - A. cellulolyticus xylA

Experiment
Protein Expression Prokaryotic cells - A. cellulolyticus xylA
Product
pANC209 (xylA) from Kazuhiko Ishikawa, Biomass Refinery Research Center, National In
Manufacturer
Kazuhiko Ishikawa, Biomass Refinery Research Center, National In

Protocol tips

Protocol tips
Protoplasts of A. cellulolyticus YP-4 were transformed with pANC209, 210, 223, 230, 231, 232 and 233 by nonhomologous integration into the host chromosomal DNA (Fujii et al. [2012]). Gene integration into prototrophic transformants was verified by genomic PCR. All cultures were grown in medium containing 20 g/l soluble starch and 5 g/l urea using the method described previously (Inoue et al. [2013]). The recombinant xylanases expressed were purified from the culture supernatant.

Publication protocol

Protoplasts of A. cellulolyticus YP-4 were transformed with pANC209, 210, 223, 230, 231, 232 and 233 by nonhomologous integration into the host chromosomal DNA (Fujii et al. [2012]). Gene integration into prototrophic transformants was verified by genomic PCR. The expression of each recombinant xylanase was carried out using the following cultures: Y209 (YP-4 transformed with pANC209; XylA), Y210 (YP-4 transformed with pANC210; XylB), Y223 (YP-4 transformed with pANC223; XylC), Y230 (YP-4 transformed with pANC230; XylD), Y231 (YP-4 transformed with pANC231; XylE), Y232 (YP-4 transformed with pANC232; XylF), Y233 (YP-4 transformed with pANC233; XylG) (Table 2). All cultures were grown in medium containing 20 g/l soluble starch and 5 g/l urea using the method described previously (Inoue et al. [2013]). The recombinant xylanases expressed were purified from the culture supernatant.

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