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===Introduction===
 
===Introduction===

Revision as of 12:31, 1 October 2023


contributed by Fudan iGEM 2023

Introduction

H. ex mtSSB is a type of mitochondrial single-stranded DNA binding protein derived from Hypsibius exemplaris . H. ex mtSSB is non-specific in binding single-stranded DNA. ssDNA is exposed by normal cellular functions like replication and transcription, as well as during genotoxic stress. DNA wrapped around H. ex mtSSB could be physically buffered against DNA damage and ensuing lethality. Under harsh conditions like desiccation heat, and radiation, H. ex mtSSB can maintain the stability of DNA through the above mentioned mechanism, thus enhance the survival rate of organisms in harsh environments[1].

Usage and Biology

We heterologously expressed H. ex mtSSB in E. coli, conferring improved resistance to desiccation and UV radiation. We also compared its desiccation resistance with that of Secretory-abundant heat soluble protein 33020 (SAHS 33020) and tested the combined desiccation resistance of both proteins in ribozyme+RBS+CDS+stem-loop module: H. ex mtSSB+SAHS 33020

Characterization

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 186
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 454
    Illegal BsaI.rc site found at 535
    Illegal SapI site found at 348


Reference

  1. ↑ Hibshman, J. D., Clark-Hachtel, C. M., Bloom, K. S., & Goldstein, B. (2023). A bacterial expression cloning screen reveals tardigrade single-stranded DNA-binding proteins as potent desicco-protectants (2023.08.21.554171). bioRxiv. https://doi.org/10.1101/2023.08.21.554171