Difference between revisions of "Part:BBa K1151001:Experience"

Latest revision as of 05:19, 2 November 2014

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1151001

User Reviews

UNIQ6701d039aeae248b-partinfo-00000000-QINU UNIQ6701d039aeae248b-partinfo-00000001-QINU

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Imperial College 2014

We attempted to use this protein as part of fusions with cellulose binding domains. However after cloning our fusions using this part we noticed that unfortunately this part is out of frame, leading to an incorrect translation and read-through of the stop-codon. This also means any subsequent fusion proteins to the C-terminal are out of frame. Although the sequence updated by the authors here corresponds to the actual DNA sequence of the part, the sequence itself is erroneous as it contains a deletion with respect to the original Heliobacter pylori NiBP (GenBank CP000012.1 bp1391851-1392033, reverse complement strand). To correct the mutation a 'C' needs to be added between bases 52-53. This should be able to be easily achieved using site-directed mutagenesis PCR.

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 <I>Imperial College 2014</I>
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-We attempted to use this protein as part of fusions with cellulose binding domains. However after cloning our fusions using this part we noticed that unfortunately this part is out of frame, leading to an incorrect translation and read-through of the stop-codon. This also means any subsequent fusion proteins to the C-terminal are out of frame. This is because there is a deletion of a base compared to the equivalent coding sequence from Heliobacter pylori (GenBank CP000012.1 bp1391851-1392033, reverse complement strand). To correct the mutation a 'C' needs to be added between bases 52-53. This should be able to be easily achieved using site-directed mutagenesis PCR.
+We attempted to use this protein as part of fusions with cellulose binding domains. However after cloning our fusions using this part we noticed that unfortunately this part is out of frame, leading to an incorrect translation and read-through of the stop-codon. This also means any subsequent fusion proteins to the C-terminal are out of frame. Although the sequence updated by the authors here corresponds to the actual DNA sequence of the part, the sequence itself is erroneous as it contains a deletion with respect to the original ''Heliobacter pylori'' NiBP (GenBank CP000012.1 bp1391851-1392033, reverse complement strand). To correct the mutation a 'C' needs to be added between bases 52-53. This should be able to be easily achieved using site-directed mutagenesis PCR.
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