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'''Figure 1.'''This is the pre-expression using E.coli BL21 at 37 ℃.<br>
 
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'''Figure 1.'''This is the pre-expression using E.coli BL21 at 37 ℃.<br>
 
 
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'''Figure 2.''' This is the pre-expression using E.coli BL21 at 16 ℃. <br>
 
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'''Figure 2.''' This is the pre-expression using E.coli BL21 at 16 ℃. <br>
 
 
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     https://static.igem.org/mediawiki/igem.org/9/94/Tjuresults42.jpg<br>
 
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'''Figure 3.''' This is the pre-expression using E.coli BL21 at 25 ℃. <br>
 
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'''Figure 3.''' This is the pre-expression using E.coli BL21 at 25 ℃. <br>
 
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===Surface display HPLC results===
 
===Surface display HPLC results===
 
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'''Figure 4.''' Relative enzyme activity of  engineering bacteria E.coli(BL21)/pET22b(+)LAP when induced at 16℃. <br>
 
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'''Figure 4.''' Relative enzyme activity of  engineering bacteria E.coli(BL21)/pET22b(+)LAP when induced at 16℃. <br>
 

Latest revision as of 07:18, 20 October 2016


LPP-OmpA

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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage

The Lpp-OmpA consists of the first nine N-terminal amino acids of major E. coli lipoprotein (Lpp) bined with a transmembrane domain (46-159 aa) from outer membrane protein A (OmpA). With the help of Lpp-OmpA, we can direct different polypeptides to the cell surface of E. coli, such as β-lactamase, cellulases, etal. Now we hope to display our PETase on the surface of E. coli by using it.

Biology

Surface expression of recombinant proteins was first described more than 30 years ago.The Lpp-OmpA consists of the first nine N-terminal amino acids of major E. coli lipoprotein (Lpp) bined with a transmembrane domain (46-159 aa) from outer membrane protein A (OmpA). We got its sequence from the genome of Escherichia coli str. K-12 substr.

Reference

[1] Ali Karami, Ali Mohamad Latifi*, and Samaneh Khodi，etal. 2014. Comparison of the Organophosphorus Hydrolase Surface Display Using InaVN and Lpp-OmpA Systems in Escherichia coli. J. Microbiol. Biotechnol. (2014),24(3), 379–385 [2] Joseph A. Francisco*, etal. 1992. Transport and anchoring of β-lactamase to the external surface of Escherichia coli.

Pre-expression

Tjuresults41.jpg
Figure 1.This is the pre-expression using E.coli BL21 at 37 ℃.

Tjuresults43.jpg
Figure 2. This is the pre-expression using E.coli BL21 at 16 ℃.

Tjuresults42.jpg
Figure 3. This is the pre-expression using E.coli BL21 at 25 ℃.

Surface display HPLC results

ProofTJU8.jpg
Figure 4. Relative enzyme activity of engineering bacteria E.coli(BL21)/pET22b(+)LAP when induced at 16℃.