Difference between revisions of "Part:BBa K2273108"

Revision as of 13:44, 25 September 2017

BlaR1 Beta-Lactam Receptor derived from Staphylococcus aureus N315 The blaR1 gene is part of the Beta-Lactam Biosensor project of [http://2017.igem.org/Team:TU_Dresden iGEM Team TU Dresden 2017 (EncaBcillus - It's a trap!)].

This gene is part of the bla operon found in Staphylococcus aureus and encodes a receptor that localizes in the inner cell membrane and can bind beta-lactam antibiotics, Uniprot [http://www.uniprot.org/uniprot/P18357]). This Part serves as a sensing unit of the beta-lactam biosensor. The BlaR1 receptor is able to bind various beta-lactam antibiotics. After binding of a respective ligand, a signal is transducer into the bacterial cytoplasm. This Protein therefore harbours transmembrane helices and a Penicillin Binding domain (amino acid 331-585

This part has been codon optimized for expression in Bacillus subtilis using the online tool provided by IDT DNA. A Ribosome Binding Site (AGGAGG) specific for translation in Bacillus subtilis has been added upstream of the gene followed by a seven nucleotide spacer. Further the part features the RFC10 prefix and suffix:

Prefix with	EcoRI, NotI, XbaI, RBS and spacer sequence	GAATTCGCGGCCGCTTCTAGAAGGAGGTGTCAAA
Suffix with	SpeI, NotI and PstI	ACTAGTAGCGGCCGCTGCAGA

Sites of restriction enzymes generating compatible overhangs are indicated by sharing one color. (EcoRI and PstI are marked in blue, NotI in green, XbaI and SpeI in red

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]

Beta-Lactam Biosensor

In this subproject, we developed a functional and complete heterologous beta-lactam biosensor in Bacillus subtilis. By the time these specified cells sense a compound of the beta-lactam family, they will respond by producing a measurable luminescence signal. We further investigated the detection spectrum of the biosensor by testing different beta-lactam antibiotics from various subclasses. For increased control and easy handling of the biosensor strain during a potential field application, we demonstrate that the encapsulation of the cells into Peptidosomes is quite advantageous.

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K2273108 short</partinfo>
 The <i>blaR1</i> gene is part of the Beta-Lactam Biosensor project of [http://2017.igem.org/Team:TU_Dresden iGEM Team TU Dresden 2017 (EncaBcillus - It's a trap!)].<br><br>
-This gene is part of the <i>bla</i> operon found in <i>Staphylococcus aureus</i> and encodes a receptor that localizes in the inner cell membrane and can bind beta-lactam antibiotics, Uniprot [http://www.uniprot.org/uniprot/P18357]).<br><br>
+This gene is part of the <i>bla</i> operon found in <i>Staphylococcus aureus</i> and encodes a receptor that localizes in the inner cell membrane and can bind beta-lactam antibiotics, Uniprot [http://www.uniprot.org/uniprot/P18357]). This Part serves as a sensing unit of the beta-lactam biosensor. The BlaR1 receptor is able to bind various beta-lactam antibiotics. After binding of a respective ligand, a signal is transducer into the bacterial cytoplasm. This Protein therefore harbours transmembrane helices and a Penicillin Binding domain (amino acid 331-585<br><br>
 This part has been codon optimized for expression in <i>Bacillus subtilis</i> using the online tool provided by IDT DNA. A Ribosome Binding Site (AGGAGG) specific for translation in <i>Bacillus subtilis</i> has been added upstream of the gene followed by a seven nucleotide spacer. Further the part features the RFC10 prefix and suffix:<br>
@@ Line 26: / Line 26: @@
 <!-- Add more about the biology of this part here
 ===Usage and Biology===
-This Part serves as a sensing unit of the beta-lactam biosensor. The BlaR1 receptor is able to bind various beta-lactam antibiotics. After binding of a respective ligand, a signal is transducer into the bacterial cytoplasm. This Protein therefore harbours transmembrane helices and a Penicillin Binding domain (amino acid 331-585).
 <!-- -->