Difference between revisions of "Part:BBa K4375017"

Revision as of 09:54, 10 October 2022

anti-CEA Nanobody Display System

This construction consists of 4 described parts. These are: J23101* (BBa_K4375003), anti-CEA-specific Nanobody, (BBa_K4375006 ), Intimin (BBa_K4375012), Myc (BBa_K4375014). They allow the bacterium to bind to the cancer cell surface, thus allowing its visualization and colonization on the tumor surface.

Usage and Biology

This construct is meant for detecting cancer cells that express CEA, carcioembrional antigen by bacterial cells. It is expressed by the bacterial cells on their surface and can be used to detect the tumor by as it causes the bacteria to bind to cancer cells. This device contains two different parts whose production is controlled by a constitutive strong promoter J23101* (BBa_K4375003). The anti-CEA Nanobody (BBa_K4375006) is at the N-terminal, tagged with a c-myc for detection. The Nanobody is fused to the Intimin part, which is a 12-stranded β-barrel protein found in bacterial adhesins (BBa_K4375012). Specific interaction can be detected by two tags: Myc (BBa_K4375014) and anti-E, or via the fluorescence of the sfGFP at the C-terminal.

Intimin and CEA nanobody conjugation scheme.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 29
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 2043
Illegal BamHI site found at 4059
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 2064
Illegal NgoMIV site found at 4080
Illegal AgeI site found at 4146
Illegal AgeI site found at 4278
Illegal AgeI site found at 4678
1000
COMPATIBLE WITH RFC[1000]

@@ Line 4: / Line 4: @@
-This construction consists of 4 described parts. These are:  J23101* (BBa_K4375003), anti-CEA-specific Nanobody, (BBa_K4375006 ),  Intimin (BBa_K4375012),  Myc (BBa_K4375014). They allow the bacterium to be expelled to the tumour surface, thus allowing its visualisation and colonisation on the tumour surface.
+This construction consists of 4 described parts. These are:  J23101* (BBa_K4375003), anti-CEA-specific Nanobody, (BBa_K4375006 ),  Intimin (BBa_K4375012),  Myc (BBa_K4375014). They allow the bacterium to bind to the cancer cell surface, thus allowing its visualization and colonization on the tumor surface.
 ==Usage and Biology==
-This construct is meant for detecting cancer cells that express CEA, carcioembrional antigen by bacterial cells. This device contains two different constructs whose production is under the control of a constitutive strong promoter J23101* (BBa_K4375003). CEA-positive cell-specific attachment is achieved by an anti-CEA-specific Nanobody, (BBa_K4375006 ). The Nanobody displayed on the bacteria outer membrane, by the N-terminal domain of a bacterial adhesion factor, Intimin, described in BBa_K4375012. Cancer cell specific interaction can be detected by two epitope tags: Myc (BBa_K4375014) and anti-E, or via the fluorescence of the policistronicly expressed sfGFP.
+This construct is meant for detecting cancer cells that express CEA, carcioembrional antigen by bacterial cells. It is expressed by the bacterial cells on their surface and can be used to detect the tumor by as it causes the bacteria to bind to cancer cells. This device contains two different parts whose production is controlled by a constitutive strong promoter J23101* (BBa_K4375003). The anti-CEA Nanobody (BBa_K4375006) is at the N-terminal, tagged with a c-myc for detection. The Nanobody is fused to the Intimin part, which is a 12-stranded β-barrel protein found in bacterial adhesins (BBa_K4375012). Specific interaction can be detected by two tags: Myc (BBa_K4375014) and anti-E, or via the fluorescence of the sfGFP at the C-terminal.
+[[File: BBa K4375017 IntiminCEA.png]]
+Intimin and CEA nanobody conjugation scheme.