pANC209 (xylA)

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	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.

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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