pBNS2-man

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To further verify the constructed plasmids, the β-mannanase gene (GeneBank Accession No. KC436314) was inserted into the SD sequence of pBNS2, and the β-mannanase activity was measured. Using primers manA-F/manA-D, the β-mannanase (named manB) gene was amplified from Bacillus pumilus Nsic-2. Next, the manB gene was subcloned into the BamHI/XhoI site of pBNS2 to generate pBNS2-man. Sequence verification of the manB gene in pBNS2-man was performed.	The recombinant plasmid was then transformed into the B. subtilis 1A751 strain.

Upstream tips
To further verify the constructed plasmids, the β-mannanase gene (GeneBank Accession No. KC436314) was inserted into the SD sequence of pBNS2, and the β-mannanase activity was measured. Using primers manA-F/manA-D, the β-mannanase (named manB) gene was amplified from Bacillus pumilus Nsic-2. Next, the manB gene was subcloned into the BamHI/XhoI site of pBNS2 to generate pBNS2-man. Sequence verification of the manB gene in pBNS2-man was performed.
Protocol tips
The recombinant plasmid was then transformed into the B. subtilis 1A751 strain.

Publication protocol

"To further verify the constructed plasmids, the β-mannanase gene (GeneBank Accession No. KC436314) was inserted into the SD sequence of pBNS2, and the β-mannanase activity was measured. Using primers manA-F/manA-D, the β-mannanase (named manB)
gene was amplified from Bacillus pumilus Nsic-2. Next, the manB gene was subcloned into the BamHI/XhoI site of pBNS2 to generate pBNS2-man. Sequence verification of the manB gene in pBNS2-man was performed. The recombinant plasmid was then transformed into the B. subtilis 1A751 strain."

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