P. xylaniclasticus strain TW1 was isolated previously from the wastes of a pineapple processing factory in Thailand.5) (link) The Escherichia coli strains αINV (Invitrogen, Thermo Fisher Scientific, Inc., Waltham, MA, USA) and ME9806 (iVEC3) (National BioResource Project (NBRP), Mishima, Japan) were used as cloning hosts while E. coli JM109 and BL21(DE3)(TOYOBO CO., LTD., Japan) were used as protein expression hosts.7) (link) The pCR2.1 plasmid (Invitrogen) was used for cloning, and the pQE30 plasmid (QIAGEN Benelux B.V., Venlo, Netherland) and pET16b plasmid (Novagen Inc., Madison, WI, USA) were used for expression of recombinant His-tagged proteins. Transformed E. coli was cultivated in LB liguid medium supplemented with ampicillin (50 µg/mL). The recombinant proteins of the full length of PxXyl43A (PxXyl43A) and the unknown function module at the C-terminus of PxXyl43A (PxXyl43A-UM) were expressed using the plasmids pET16b-PxXyl43A and pQE30-PxXyl43A-UM, respectively.
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Recombinant Protein Expression of PxXyl43A
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- Cloning hosts: αINV (Invitrogen), ME9806 (iVEC3)
- Protein expression hosts: E. coli JM109, BL21(DE3)
- Plasmids: pCR2.1, pQE30, pET16b
- Expression of recombinant His-tagged proteins: PxXyl43A and PxXyl43A-UM
- Cultivation of transformed E. coli in LB liquid medium supplemented with ampicillin (50 µg/mL)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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