Difference between revisions of "Part:BBa K2918003"
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<partinfo>BBa_K2918003 short</partinfo> | <partinfo>BBa_K2918003 short</partinfo> | ||
| − | DSB of | + | DSB of Φ29 bacteriophage |
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| + | ===Overview=== | ||
| + | |||
| + | The replication of DNA and its conversion into functional proteins are vital processes in all living systems. DNA is copied during the replication process. The bacteriophage Φ29 contains a DNA replication machinery which replicates the linear plasmid by itself. This process is called orthogonal replication and can be beneficially used. The desired gene can be expressed in other hosts without interfering with the genome of its host. Our Sci-Phi 29 tool is based on the Φ29 DNA replication system and its four proteins. The terminal proteins (TP) cap the linear DNA, protect the linear DNA and are the starting point for the Φ29 DNA polymerase (DNAP/p2). DNAP binds to the TP and replicates the DNA. During the replication, single and double stranded DNA is protected by respectively single stranded DNA binding proteins (SSB/p5) and double stranded DNA binding proteins (DSB/p6). | ||
| + | |||
| + | ===Strain Construction=== | ||
| + | |||
| + | <b>Aim:</b> To clone the promoter in a level 0 MoClo backbone <html>(<a target=”_blank” href=”https://www.addgene.org/47984/”>pICH41233</a>)</html>. <br> | ||
| + | <b>Procedure:</b> The DNA sequence of the part was synthesized by IDT with flanking Bpi1 sites and respective MoClo compatible promoter overhangs. The cloning protocol can be found in the MoClo section below. | ||
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Revision as of 21:40, 3 October 2019
Φ29 Double Stranded Binding Protein (DSB/p6)
DSB of Φ29 bacteriophage
Overview
The replication of DNA and its conversion into functional proteins are vital processes in all living systems. DNA is copied during the replication process. The bacteriophage Φ29 contains a DNA replication machinery which replicates the linear plasmid by itself. This process is called orthogonal replication and can be beneficially used. The desired gene can be expressed in other hosts without interfering with the genome of its host. Our Sci-Phi 29 tool is based on the Φ29 DNA replication system and its four proteins. The terminal proteins (TP) cap the linear DNA, protect the linear DNA and are the starting point for the Φ29 DNA polymerase (DNAP/p2). DNAP binds to the TP and replicates the DNA. During the replication, single and double stranded DNA is protected by respectively single stranded DNA binding proteins (SSB/p5) and double stranded DNA binding proteins (DSB/p6).
Strain Construction
Aim: To clone the promoter in a level 0 MoClo backbone (pICH41233).
Procedure: The DNA sequence of the part was synthesized by IDT with flanking Bpi1 sites and respective MoClo compatible promoter overhangs. The cloning protocol can be found in the MoClo section below.
Sequence and Features
- 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 168
- 1000COMPATIBLE WITH RFC[1000]