Translational_Unit

Part:BBa_K4030007

Designed by: Nixue Song   Group: iGEM21_Shanghai_United_HS   (2021-10-12)
Revision as of 01:40, 20 October 2021 by Hrr1225 (Talk | contribs)


J23101-OmpA-arab-TT

Profile

Name: J23101-OmpA-arab-TT

Base Pairs: 3611bp

Origin: E. coli, synthetic

Properties: Arabinose expression system

Usage and Biology

Dental caries is a common disease. It not only directly affects human oral health, but also often causes adverse symptoms in other parts of the body. Global disease statistics in 2016 show that the incidence of dental caries in the population is ranking second among common diseases. Studies have shown that the formation of dental plaque is the result of the joint action of a variety of bacteria, including Streptococcus mutans, Lactobacillus, Actinomycetes, etc. Phage lyase ClyR (combined from different bacteriophage lytic enzymes) has a broad bactericidal spectrum, especially the only one reported to be extremely strong against Streptococcus mutans and Streptococcus mulberry. The enzyme is promising to kill these two kinds of streptococci are the main cause of dental caries.

Figure 1. Schematic diagram of ClyR in the prevention or treatment of dental caries..

Construct design

Arab is connected with OpmA and this sequence is inserted into plasmid pUC57_mini (Figure 2 and 3).

Figure 2. J23101-OmpA-arab-TT box..
Figure 3. Schematic map of J23101-OmpA-arab-TT plasmid..

The profiles of every basic part are as follows:

BBa_K4030001

Name: arab

Base Pairs: 1479bp

Origin: Bacillus pumilus

Properties: Secreting arabinosidase to hydrolyze polysaccharides into monosaccharides.

Usage and Biology

BBa_K4030001 is a coding sequence of arab, which is an enzyme used to hydrolyze arabinose from arabinoxylans, a carbohydrate that appears normally in foods.

BBa_K4030000

Name: OmpA

Base Pairs: 63bp

Origin: Escherichia coli

Properties: Outer membrane protein A

Usage and Biology

Outer membrane protein A (OmpA) is a major protein in the Escherichia coli outer membrane.


Experimental approach

Group 1 Plasmid puc57-kan-mini-J23101-OmpA-araB-TT (Plasmid A) was transformed to E. coli Nissle 1917 by electroporation. We tried to monitor the concentration of the reducing sugar arabinose and thus access the activity of AraB. The concentration of reducing sugar was determined by the DNS kit and the A540 were recorded accordingly.














Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 745
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 745
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 745
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 745
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 745
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None