Difference between revisions of "Part:BBa K4879005"
Sauron 3791 (Talk | contribs) |
Sauron 3791 (Talk | contribs) |
||
| Line 9: | Line 9: | ||
RAD52 is a well-documented protein that helps in facilitating homologous recombination and subsequent DNA repair; found to be well-conserved across many species. It assists in the formation of a nucleoprotein filament on the broken single-stranded DNA (ssDNA) and thus facilitates strand invasion, the first step in homologous DNA repair. | RAD52 is a well-documented protein that helps in facilitating homologous recombination and subsequent DNA repair; found to be well-conserved across many species. It assists in the formation of a nucleoprotein filament on the broken single-stranded DNA (ssDNA) and thus facilitates strand invasion, the first step in homologous DNA repair. | ||
| − | Wild-type <i>Y.lipolytica</i> has a very low probability of homologous recombination, which was detrimental to the deletion attempts we made to delete <i>faa1</i> and <i>alk2</i> genes from our chassis. As ScRAD52 enhances this probability to as high as | + | Wild-type <i>Y.lipolytica</i> has a very low probability of homologous recombination (about 0-36%), which was detrimental to the deletion attempts we made to delete <i>faa1</i> and <i>alk2</i> genes from our chassis. As ScRAD52 enhances this probability to as high as 95%, we inserted the [https://parts.igem.org/Part:BBa_K4879011 (ScRAD52 expression construct)] into our organism. |
<html><img src="https://static.igem.wiki/teams/4879/wiki/bba-k4879005-scrad52.png"width="250"height="250"></html> | <html><img src="https://static.igem.wiki/teams/4879/wiki/bba-k4879005-scrad52.png"width="250"height="250"></html> | ||
Latest revision as of 09:19, 11 October 2023
Yarrowia lipolytica ScRAD52 gene
RAD52 coding sequence from Saccharomyces cerevisiae, codon optimized for Yarrowia lipolytica. [1]
Usage and Biology
RAD52 is a well-documented protein that helps in facilitating homologous recombination and subsequent DNA repair; found to be well-conserved across many species. It assists in the formation of a nucleoprotein filament on the broken single-stranded DNA (ssDNA) and thus facilitates strand invasion, the first step in homologous DNA repair.
Wild-type Y.lipolytica has a very low probability of homologous recombination (about 0-36%), which was detrimental to the deletion attempts we made to delete faa1 and alk2 genes from our chassis. As ScRAD52 enhances this probability to as high as 95%, we inserted the (ScRAD52 expression construct) into our organism.
The protein structure of ScRAD52, as predicted by AlphaFold.
Design
The transcriptional unit for CvFAP (BBa_K4879011) is designed with the coding sequence flanked by the TEF1 promoter (BBa_K2983053) upstream and the XPR2 terminator (BBa_K3629004) downstream.
References
1. Qingchun Ji, Jie Mai, Ying Ding, Yongjun Wei, Rodrigo Ledesma-Amaro, Xiao-Jun Ji, Improving the homologous recombination efficiency of Yarrowia lipolytica by grafting heterologous component from Saccharomyces cerevisiae, Metabolic Engineering Communications, Volume 11, 2020, e00152, ISSN 2214-0301, https://doi.org/10.1016/j.mec.2020.e00152.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 403
Illegal BglII site found at 533
Illegal XhoI site found at 1223 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1207
- 1000COMPATIBLE WITH RFC[1000]
References
1. Qingchun Ji, Jie Mai, Ying Ding, Yongjun Wei, Rodrigo Ledesma-Amaro, Xiao-Jun Ji. Improving the homologous recombination efficiency of Yarrowia lipolytica by grafting heterologous component from Saccharomyces cerevisiae, Metabolic Engineering Communications, Volume 11, 2020, e00152, ISSN 2214-0301 https://doi.org/10.1016/j.mec.2020.e00152