Difference between revisions of "Part:BBa K4879005"

 
 
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<partinfo>BBa_K4879005 short</partinfo>
 
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RAD52 coding sequence from <i>Saccharomyces cerevisiae</i>, codon optimized for <i>Yarrowia lipolytica</i>. [1]
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===Usage and Biology===
 
===Usage and Biology===
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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.
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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.
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<html><img src="https://static.igem.wiki/teams/4879/wiki/bba-k4879005-scrad52.png"width="250"height="250"></html>
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The protein structure of ScRAD52, as predicted by AlphaFold.
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===Design===
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The transcriptional unit for CvFAP [https://parts.igem.org/Part:BBa_K4879011 (BBa_K4879011)] is designed with the coding sequence flanked by the TEF1 promoter [https://parts.igem.org/Part:BBa_K2983053 (BBa_K2983053)] upstream and the XPR2 terminator [https://parts.igem.org/Part:BBa_K3629004 (BBa_K3629004)] downstream.
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===References===
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1. Qingchun Ji, Jie Mai, Ying Ding, Yongjun Wei, Rodrigo Ledesma-Amaro, Xiao-Jun Ji,
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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,
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https://doi.org/10.1016/j.mec.2020.e00152.
  
 
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<partinfo>BBa_K4879005 parameters</partinfo>
 
<partinfo>BBa_K4879005 parameters</partinfo>
 
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===References===
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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
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https://doi.org/10.1016/j.mec.2020.e00152

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 403
    Illegal BglII site found at 533
    Illegal XhoI site found at 1223
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1207
  • 1000
    COMPATIBLE 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