Part:BBa_K2375000

PurA Gene

PurA gene extracted from E. coli. Golden Gate Synthesis was used to remove the illegal EcoRI site.

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 AgeI site found at 388
  Illegal AgeI site found at 751
  Illegal AgeI site found at 1246
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 1210
  

Contribution From NNU-China 2021

Group: iGEM Team NNU-China 2021

Author: Yan Xu

Summary: Charactering its function on growth and AMPs function

Characterization from iGEM21-NNU-China

        The purA gene was registered in 2017 as (BBa_K2375000). The purA is the critical gene in the purine metabolism in E.coli, which can impact the protein biosynthesis. Using our enzyme-constrained model, it is predicted that reduced purA expression is beneficial to improve the production of antimicrobial peptides (AMPs). Here, we characterized the effect of purA on cell growth and production of AMPs (Fig.4). We deleted the purA gene from the genome of BL21 (DE3), yielding the strain of △purA. Compared with the BL21(DE3), △purA showed a better growth. Moreover, the production of SFa and SMAP was increased by 150% and 63% than that of BL21 (DE3). These results provide references for future iGEM team to enhance the production of AMPs or hard-to-express proteins.

Fig.1 A. The growth of BL21(DE3) and △purA strain. B. The sFa and SMAP expression in the BL21 (DE3) and △purA strain.

Reference

1. Yoshitoshi O, Yukiho I, Naotake O, Shigeki M. Autoregulation of the dnaA-dnaN operon and effects of dnaA protein levels on replication initiation in bacillus subtilis. Journal of Bacteriology. 2001; 183(13): 3833-3841.