Difference between revisions of "Part:BBa K133038"

Latest revision as of 02:42, 30 October 2008

T7-CF-RGD-Histop

This part was used in a vector with T7 promotor. The vector with the part inserted was then transformed into bacterial cells with lambdaDE3 phage DNA, which enabled the expression of the protein, induced by IPTG.

Chimeric fusion protein, which retains the central, most antigenic segment of H. pylori (hypervariable region) flagellin FlaA, while replacing the N- and C-terminal segments (176 aa from the N-terminal and 99 aa from the C-terminal) by the FliC flagellin from E. coli, which is responsible for the activation of TLR5. Thus, combining the right parts of flagellin of E. coli with that of H. pylori enables activation of innate immunity - a characteristic that flagellin of H. pylori alone does not posess - while presenting the antigen of H. pylori for antibody production.

The protein was to be produced, therefore we added RGD His stop at the end of the construct.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 1207
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 764
Illegal BamHI site found at 1275
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 301
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1558

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 This part was used in a vector with T7 promotor. The vector with the part inserted was then transformed into bacterial cells with lambdaDE3 phage DNA, which enabled the expression of the protein, induced by IPTG.
-Chimeric fusion protein, which retains the central, most antigenic segment of H. pylori (hypervariable region) flagellin FlaA, while replacing the N- and C-terminal segment by the FliC flagellin from E. coli, which is responsible for the activation of TLR5 (176 aa from the N-terminal and 99 aa from the C-terminal). Thus, combining the right parts of flagellin of ''E. coli'' with that of ''H. pylori'' enables activation of innate immunity - a characteristic that flagellin of ''H. pylori'' alone does not posess - while presenting the antigen of ''H. pylori'' for antibody production.
+Chimeric fusion protein, which retains the central, most antigenic segment of H. pylori (hypervariable region) flagellin FlaA, while replacing the N- and C-terminal segments (176 aa from the N-terminal and 99 aa from the C-terminal) by the FliC flagellin from E. coli, which is responsible for the activation of TLR5. Thus, combining the right parts of flagellin of ''E. coli'' with that of ''H. pylori'' enables activation of innate immunity - a characteristic that flagellin of ''H. pylori'' alone does not posess - while presenting the antigen of ''H. pylori'' for antibody production.
 The protein was to be produced, therefore we added RGD His stop at the end of the construct.