pET30a(+)-MSP2

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	The recombined plasmids were transformed into E. coli BL21(DE3) and induced for expressing with 0.2 mM isopropyl-1-thio-β-d-galactopyranoside (IPTG) for 7 h.

Protocol tips
The recombined plasmids were transformed into E. coli BL21(DE3) and induced for expressing with 0.2 mM isopropyl-1-thio-β-d-galactopyranoside (IPTG) for 7 h.

Publication protocol

The ligation products were transformed into Escherichia coli DH5α, positive transformants with the appropriate insert were screened on medium supplemented with 50 μl/ml kanamycin, and the recombinant plasmids were identified by PCR amplification with the msp2-F and msp2-R primers and DNA sequencing. The recombined plasmids were transformed into E. coli BL21(DE3) and induced for expressing with 0.2 mM isopropyl-1-thio-β-d-galactopyranoside (IPTG) for 7 h. Cells were centrifuged (5,000 rpm for 10 min) at 4°C and resuspended in 50 mM phosphate buffer (pH 7.4). The cells were lysed by sonication, and the cellular debris was removed by centrifugation at 8,000 rpm for 15 min at 4°C. The water-soluble fraction containing all potential hexa-His tag proteins was filtered (0.2 μm filter), and purification of the MSP2 recombinant protein was performed by using Ni-NTA (nitrilotriacetic acid) His-bind resin as described by the manufacturer (Novagen, Madison, WI).

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