Difference between revisions of "Part:BBa K2752012"
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A plasmid containing a urate oxidase sequence was introduced into the BL21 strain, and IPTG was used as an inducer to induce expression of urate oxidase. After expression, the cells were disrupted by sonication and the supernatant was taken. The supernatant (extracellular protein) of the first centrifugation and the supernatant (intracellular protein) after sonication were subjected to polyacrylamide gel electrophoresis. The electrophoresis results are shown in Figure 1. | A plasmid containing a urate oxidase sequence was introduced into the BL21 strain, and IPTG was used as an inducer to induce expression of urate oxidase. After expression, the cells were disrupted by sonication and the supernatant was taken. The supernatant (extracellular protein) of the first centrifugation and the supernatant (intracellular protein) after sonication were subjected to polyacrylamide gel electrophoresis. The electrophoresis results are shown in Figure 1. | ||
| + | |||
| + | [[File:Figure 1-uox.png|400px|thumb|Figure 1. '''Polyacrylamide gel electrophoresis result, red arrow indicates the band urate oxidase.''' | ||
| + | BP: Negative control (blank) precipitation; | ||
| + | BS: Negative control (blank) supernatant; | ||
| + | PD: Precipitation after disruption of cells; | ||
| + | SD: Supernatant after disruption of cells; | ||
| + | M: Marker; | ||
| + | ]] | ||
| + | |||
Revision as of 16:13, 17 October 2018
Uox
This urate oxidase is novel mammalian uricase protein, containing some human uricase chimeric protein having an amino acid sequence of the uricase activity of mammalian origin and mutant proteins. The amino acid sequence of the 241-304th of the urate oxidase is derived from human, and the amino acid sequence of the 1-240th is derived from the amino acid sequence of the uricase of pigs and dogs.
Sequence and Features
Characterization
A plasmid containing a urate oxidase sequence was introduced into the BL21 strain, and IPTG was used as an inducer to induce expression of urate oxidase. After expression, the cells were disrupted by sonication and the supernatant was taken. The supernatant (extracellular protein) of the first centrifugation and the supernatant (intracellular protein) after sonication were subjected to polyacrylamide gel electrophoresis. The electrophoresis results are shown in Figure 1.
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 55
Illegal AgeI site found at 406
Illegal AgeI site found at 877 - 1000COMPATIBLE WITH RFC[1000]