Difference between revisions of "Part:BBa K3078004"
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To verify the construction of pVE-&#946;-1,3-glucanase(pVE-&#946;-GA) which we generated, the digestion by SalI/EcoRV was performed by a standard protocol following agarose gel electrophoresis (Figure 1).
 
To verify the construction of pVE-&#946;-1,3-glucanase(pVE-&#946;-GA) which we generated, the digestion by SalI/EcoRV was performed by a standard protocol following agarose gel electrophoresis (Figure 1).
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This year our team registered the superfolder GFP designed by Overkamp W et al with a BBa_K2541400 (called sfGFP_optimism). Compared with superfolder GFP (BBa_I746916), sfGFP_optimism (BBa_K2541400) is BbsI restriction site free, and the BbsI restriction endonuclease is an economical and efficient enzyme used in Golden Gate assembly, so sfGFP_optimism can be used in Golden Gate assembly to achieve efficient and rapid assembly of gene fragments.
 
 
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[[File:Jilin_China-sfGFP-1.0.png|center|Jilin_China-sfGFP-1.0]]
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[[File:B13-1.png|center|B13-1]]
 
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Figure 1. Expression of three types of sfGFP(BBa_I746916, BBa_K2541401, BBa_K2541400), cultivated overnight.
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Figure 1. Digestion and agarose gel electrophoresis of pVE-&#946;-GA.
 
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Revision as of 01:37, 21 October 2019


β-1,3-glucanase

β-1,3-glucanase protein coding region. β-1,3-glucanase can degrade biofilm.

1. Usage and Biology

β-1,3-glucan is one of the primary components in C. albicans biofilm EPS, which is important for Candida biofilm formation and resistance to stresses. The enzyme β-1,3-glucanase, form Cellulosimicrobium cellulans, can degrade β-1,3-glucan. Therefore, this year, we decided use β-1,3-glucanase to disrupt the Candida biofilm matrix and increase the effect of the antimicrobial drug.


2. Characterization

We characterised β-1,3-glucanase by cloning it into pVE vector. Moreover, an signal peptide were added.

To verify the construction of pVE-β-1,3-glucanase(pVE-β-GA) which we generated, the digestion by SalI/EcoRV was performed by a standard protocol following agarose gel electrophoresis (Figure 1).

B13-1

Figure 1. Digestion and agarose gel electrophoresis of pVE-β-GA.











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 274
    Illegal NgoMIV site found at 483
    Illegal NgoMIV site found at 622
  • 1000
    COMPATIBLE WITH RFC[1000]