Part:BBa_K3580200:Experience

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Applications of BBa_K3580200

Expression and purification of alanine racemase

In our project, we inserted this part between bacteriophage T5 promoter for E. coli RNA polymerase and lambda t0 terminator of pCA24N(-gfp) planmid. This plasmid was transformed into the BL21(DE3) star strain. As this vector is chloramphenicol resistant, we always cultured the bacteria in media containing chloramphenicol. After expression induction by IPTG, the BL21 cells were collected by centrifugation, then sonicated and His-Tag purification was performed. (wikiマテメソハイパーリンク) The His-Tag purified Alr was confirmed with SDS-PAGE(Fig.1). The meanings of each abbreviation are given in Table 1. The concentration of purified racemase was quantified with the Bradford method.

Fig.1 SDS-PAGEs of purified E. coli BL21(DE3)star containing BBa_K3580200

Table.1 Each meaning of abbriviation

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