Difference between revisions of "Part:BBa K3733008"
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<p>The plasmid we designed consists of T7 promoter, lac operator, RBS, HyLα coding sequence, His tag and T7 terminator, which are arranged in an order on a pET28 backbone. We aim to induce the transcription of the downstream HyLα by adding the IPTG to initiate the expression. The protein will then be purified and antibacterial activity was tested by culturing the <i>Salmonella enterica subsp. Enterica</i> (<i>ex</i> Kauffmann and Edwards) Le Minor and Popoff serovar Typhimurium with our purified HyLα and measuring the OD<sub>600</sub> to reflect the bacteria growing condition. This part has undergone codon optimization based on the bias of <i>Escherichia coli</i>. </p> | <p>The plasmid we designed consists of T7 promoter, lac operator, RBS, HyLα coding sequence, His tag and T7 terminator, which are arranged in an order on a pET28 backbone. We aim to induce the transcription of the downstream HyLα by adding the IPTG to initiate the expression. The protein will then be purified and antibacterial activity was tested by culturing the <i>Salmonella enterica subsp. Enterica</i> (<i>ex</i> Kauffmann and Edwards) Le Minor and Popoff serovar Typhimurium with our purified HyLα and measuring the OD<sub>600</sub> to reflect the bacteria growing condition. This part has undergone codon optimization based on the bias of <i>Escherichia coli</i>. </p> | ||
| + | ===Materials and Method=== | ||
| + | |||
| + | ===Results=== | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
Revision as of 14:23, 20 October 2021
HyLα: Hybrid peptide of LL-37 and LTA
LL-37 and LTA are both affective antimicrobial peptides (AMPs). HyLα successfully combine those two AMPs by connecting the C-terminate of LTA with the N-terminate of LL-37 through GS-linker. It could neutralize Lipopolysaccharides (LPS), thus effectively blocking the downstream inflammation pathway.
Design
The plasmid we designed consists of T7 promoter, lac operator, RBS, HyLα coding sequence, His tag and T7 terminator, which are arranged in an order on a pET28 backbone. We aim to induce the transcription of the downstream HyLα by adding the IPTG to initiate the expression. The protein will then be purified and antibacterial activity was tested by culturing the Salmonella enterica subsp. Enterica (ex Kauffmann and Edwards) Le Minor and Popoff serovar Typhimurium with our purified HyLα and measuring the OD600 to reflect the bacteria growing condition. This part has undergone codon optimization based on the bias of Escherichia coli.
Materials and Method
Results
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]