Difference between revisions of "Part:BBa K3790004"
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Since the length of the albA1 fragment is less than 500bp, we chose to synthesize the sequence ourselves by Oligo assembly using Phanta polymerase. We obtained the sequence information from NCBI and designed synthetic primers for synthesis | Since the length of the albA1 fragment is less than 500bp, we chose to synthesize the sequence ourselves by Oligo assembly using Phanta polymerase. We obtained the sequence information from NCBI and designed synthetic primers for synthesis | ||
| − | [[File:T--Fudan--Oligo assembly by Taq polymerase.jpg|thumb| '''Figure 1. Oligo assembly by PCR.''' xxxxxx polymerase]] | + | [[File:T--Fudan--Oligo assembly by Taq polymerase.jpg|thumb|left|400px| '''Figure 1. Oligo assembly by PCR.''' xxxxxx polymerase]] |
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--albA1-S.ssb-E.ssb.jpg|600px|thumb| '''Figure 2. Assembled DNA binding proteins, before Sanger sequencing.''' T--Fudan--albA1-S.ssb-E.ssb ]] | + | [[File:T--Fudan--albA1-S.ssb-E.ssb.jpg|600px|thumb|left| '''Figure 2. Assembled DNA binding proteins, before Sanger sequencing.''' T--Fudan--albA1-S.ssb-E.ssb ]] |
Revision as of 03:19, 21 October 2021
albA1
Introduction
albA1 is a double-stranded binding protein from Sulfolobus solfataricus (the same species as where Sso7d from), which is close to Sso7d in length and structure. We speculate that it may have a similar function to enhance DNA polymerase activity as Sso7d.
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. And, albA1 is a double-stranded binding protein from the same species as Sso7d, Sulfolobus solfataricus, which is close to Sso7d in length and structure[1]. We speculate that albA1 may be related to Sso7d, and it may have a similar function to enhance DNA polymerase activity as Sso7d[2]. 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. 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
Since the length of the albA1 fragment is less than 500bp, we chose to synthesize the sequence ourselves by Oligo assembly using Phanta polymerase. We obtained the sequence information from NCBI and designed synthetic primers for synthesis
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.
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]
we get this sequence from ncbi.( https://www.ncbi.nlm.nih.gov/nuccore/NC_002754.1?report=fasta&from=816731&to=817024&strand=true )
Reference
- ↑ Kalichuk, V., Béhar, G., Renodon-Cornière, A. et al. The archaeal “7 kDa DNA-binding” proteins: extended characterization of an old gifted family. Sci Rep 6, 37274 (2016). https://doi.org/10.1038/srep37274
- ↑ Cao S-C, Qiu L-Z. Study of DNA binding protein DbpA affecting the performance of DNA polymerase[J]. Journal of Fudan:Natural Science Edition, 2015, 54(4):469-477.