Difference between revisions of "Part:BBa K3790002"
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The length of albA1 DNA was 288 bp, which is approximately 300 bp after adding homology arms to both ends for PCR cloning. We isolated the DNA of interest by gel extraction for subsequent reactions. | The length of albA1 DNA was 288 bp, which is approximately 300 bp after adding homology arms to both ends for PCR cloning. We isolated the DNA of interest by gel extraction for subsequent reactions. | ||
− | [[File:T--Fudan-- | + | [[File:T--Fudan--DbpA-sso10b-sso7d.jpg|600px|thumb|none| '''Figure 2. Assembled DNA binding proteins, albA1, S.ssb, E.ssb.''' 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. After PCR cloning, several bacterial clones were picked, grew into cultures and sent for Sanger sequencing. Then, we verified the sequencing results, and used the correct ones for further experiments.]] |
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===Reference=== | ===Reference=== |
Revision as of 16:09, 21 October 2021
Sso7d
Introduction
Sso7d is a double-stranded binding protein that is linked to DNA polymerase A or DNA polymerase B to produce a fusion protein with higher synthetic efficiency compared to wild-type DNA polymerase.
Usage and Biology
Sso7d is a double-stranded binding protein that is linked to DNA polymerase A or DNA polymerase B to produce a fusion protein with higher synthetic efficiency compared to wild-type DNA polymerase. The Bst Pol selected for this experiment was DNA polymerase Ⅰ, and no previous studies have focused on whether double-stranded binding proteins can enhance the activity of DNA polymerase Ⅰ[1]. However, we ventured to guess that fusing a double-stranded binding protein could enhance the related activity of DNA polymerase Ⅰ, and performed the following experiments.
Experimental Results
![](/wiki/images/6/6d/T--Fudan--Oligo_assembly_by_Taq_polymerase.jpg)
The length of albA1 DNA was 288 bp, which is approximately 300 bp after adding homology arms to both ends for PCR cloning. We isolated the DNA of interest by gel extraction for subsequent reactions.
![](/wiki/images/6/61/T--Fudan--DbpA-sso10b-sso7d.jpg)
Reference
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]
- ↑ Wang Y, Prosen DE, Mei L, Sullivan JC, Finney M, Vander Horn PB. A novel strategy to engineer DNA polymerases for enhanced processivity and improved performance in vitro. Nucleic Acids Res. 2004 Feb 18;32(3):1197-207. doi: 10.1093/nar/gkh271. PMID: 14973201; PMCID: PMC373405.