Difference between revisions of "Part:BBa K523009"

Revision as of 12:14, 10 August 2011

Exoglucanase for BioSandwich

This is a modified version of BBa_K118022 for use with the BioSandwich protocol of Edinburgh 2011.

BioSandwich assembly uses an in-frame BglII site (formed by the last 2 bases of the RFC10 prefix and the first 4 bases at the start of this BioBrick) and an in-frame SpeI site (present in the RFC10 suffix). Because the SpeI site is out of frame, two bases ("TC") have been added at the end. When combined with the T at the start of the RFC10 suffix, they code for a serine residue and correct the frameshift.

The signal peptide of the exoglucanase (amino acids 1 - 42) has been removed, since this part is intended for fusion to INP or other display proteins.

A non-synonymous change (see figure) was accidentally introduced, probably by PCR. An aspartic acid residue has become a glycine.

Usage and Biology

This is an exoglucanase from Cellulomonas fimi. It degrades cellulose by chewing at the end of a cellulose chain to produce cellobiose sugar.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NotI site found at 405
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 38
Illegal NgoMIV site found at 411
Illegal NgoMIV site found at 913
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 458
Illegal SapI.rc site found at 541

@@ Line 8: / Line 8: @@
 The signal peptide of the exoglucanase (amino acids 1 - 42) has been removed, since this part is intended for fusion to INP or other display proteins.
-A non-synonymous change (see figure) was accidentally introduced, probably by PCR.
+A non-synonymous change (see figure) was accidentally introduced, probably by PCR. An aspartic acid residue has become a glycine.
 ===Usage and Biology===