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The insert sequence is available at GenBank under accession number 1288449.
These results indicate that the insert sequence is required for the full enolase activity and may constitute the protective antigenic epitope in parasite enolase.
Sequencing and alignment of insert sequence with NCBI data base revealed; the insert shared 100%% homology with tannase gene of S. lugdunensis.
Iterative database searches starting from a domain insert sequence in bacterial β-glucosidases reveals the presence of a conserved domain shared by a wide variety of bacterial and eukaryotic proteins.
The recombinant plasmid constructs were then purified and the insert sequence confirmed by PCR and sequencing.
Clones were tested for correct insert sequence and orientation using the M13 forward and reverse primers.
Unique insert sequences were identified within each genetic selection with few clones containing the same insert sequence.
7 8 contained the shorter insert sequence 5'-ATC GGT ATT GGC TTA GCA CTG GCA ATC GGT CTT GCA.
Therefore, we went back and sequenced several more clones containing the insert sequence with no different result.
Of particular interest was the enrichment of a single insert sequence in 55 independent 5-FU resistant clones of the 163 clones analysed.
Sequencing was finished when an overall sequence quality of phred40 for the insert sequence and the vector-insert transition was achieved.
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