Difference between revisions of "Part:BBa K3790153"
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<partinfo>BBa_K3790153 short</partinfo> | <partinfo>BBa_K3790153 short</partinfo> | ||
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===Introduction=== | ===Introduction=== | ||
[[File:T--Fudan--ccic8-transparent-logo.png|100px|right|2021 Fudan]] | [[File:T--Fudan--ccic8-transparent-logo.png|100px|right|2021 Fudan]] | ||
| − | The fusion protein obtained by linking the C-terminus of Bst to the N- | + | The fusion protein obtained by linking the C-terminus of Bst to the N-terminus of albA1 via a (G2S)3 linker. |
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===Experimental Results=== | ===Experimental Results=== | ||
The length of Bst-(G2S)3-albA1 DNA was 2082 bp, which is approximately 2100 bp after adding homology arms to both ends for PCR cloning. We isolated the target DNA by gel extraction for subsequent reactions. | The length of Bst-(G2S)3-albA1 DNA was 2082 bp, which is approximately 2100 bp after adding homology arms to both ends for PCR cloning. We isolated the target DNA by gel extraction for subsequent reactions. | ||
| − | [[File:T--Fudan--bst-11.jpg|600px|thumb|none|'''Figure 1. PCR assembled Bst fusions.''' The first lane was loaded with DNA ladder, sizes were marked on the image. The brightest band of 750 bp was about 100 ng, and other bands about 50 ng. Lanes with correct sized amplified DNA were labeled. The | + | [[File:T--Fudan--bst-11.jpg|600px|thumb|none|'''Figure 1. PCR assembled Bst fusions.''' The first lane was loaded with DNA ladder, sizes were marked on the image. The brightest band of 750 bp was about 100 ng, and other bands about 50 ng. Lanes with correct sized amplified DNA were labeled. The ninth lane (lane-3) was loaded with Bst-(G2S)3-albA1 DNA.]] |
After ClonExpress homologous recombination reaction, Fast-T1 competent bacteria were transformed with reactants, and spread onto LB plates with antibiotics. We picked colony after 16 h culturing plates at 37℃. We further grow clones in 2 ml LB liquid medium with antibiotics, 37℃ shaking overnight. | After ClonExpress homologous recombination reaction, Fast-T1 competent bacteria were transformed with reactants, and spread onto LB plates with antibiotics. We picked colony after 16 h culturing plates at 37℃. We further grow clones in 2 ml LB liquid medium with antibiotics, 37℃ shaking overnight. | ||
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<span class='h3bb'>'''Sequence and Features'''</span> | <span class='h3bb'>'''Sequence and Features'''</span> | ||
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<partinfo>BBa_K3790153 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3790153 SequenceAndFeatures</partinfo> | ||
Latest revision as of 20:40, 21 October 2021
Bst-(G2S)3-albA1
Contents
Introduction
The fusion protein obtained by linking the C-terminus of Bst to the N-terminus of albA1 via a (G2S)3 linker.
Experimental Results
The length of Bst-(G2S)3-albA1 DNA was 2082 bp, which is approximately 2100 bp after adding homology arms to both ends for PCR cloning. We isolated the target DNA by gel extraction for subsequent reactions.
Figure 1. PCR assembled Bst fusions. The first lane was loaded with DNA ladder, sizes were marked on the image. The brightest band of 750 bp was about 100 ng, and other bands about 50 ng. Lanes with correct sized amplified DNA were labeled. The ninth lane (lane-3) was loaded with Bst-(G2S)3-albA1 DNA.
After ClonExpress homologous recombination reaction, Fast-T1 competent bacteria were transformed with reactants, and spread onto LB plates with antibiotics. We picked colony after 16 h culturing plates at 37℃. We further grow clones in 2 ml LB liquid medium with antibiotics, 37℃ shaking overnight.
Mini-prep was performed the following day, and the resulting plasmids were sent to Sanger sequencing.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]