Protein expression vector pET28a-mCherry-CNA35 was obtained from Addgene (#61607). Transformed Escherichia coli (BL21) was cultured in 400 ml LB, and induced protein expression with isopropyl β-D-1-tthiogalactopyranoside for 20 hr at 25℃. Cultured bacteria were spun down, and collected bacteria pellet was sonicated. The lysate was centrifuged, and the resulting supernatant was run through a column filled with N-NTA His-band resin (Millipore). Bound protein was eluted and then dialyzed for overnight in PBS at 4℃. Endotoxin was removed by endotoxin removal columns (88274; Thermo) following with manufacturer’s protocol.
Pet28a mcherry cna35
Manufactured by Addgene
PET28a-mCherry-CNA35 is a plasmid vector that contains the mCherry fluorescent protein gene fused to the CNA35 protein. The mCherry-CNA35 fusion is under the control of a T7 promoter. This plasmid can be used for recombinant protein expression in E. coli.
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2 protocols using pet28a mcherry cna35
1
Purification of mCherry-CNA35 Fusion Protein
2
Purification of mCherry-CNA35 Fusion Protein
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Protein expression vector pET28a-mCherry-CNA35 was obtained from Addgene (#61607). Transformed E.coli (BL21) was cultured in 400 ml LB, and induced protein expression with isopropyl β-D-1-tthiogalactopyranoside for 20 hours at 25 . Cultured bacteria were spun down, and collected bacteria pellet was sonicated. The lysate was centrifuged and the resulting supernatant was run through a column filled with N-NTA His-band resin (Millipore). Bound protein was eluted and then dialyzed for overnight in PBS at 4 . Endotoxin was removed by endotoxin removal columns (88274; Thermo) following with manufacturer's protocol.
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