The engineered donor sequence is integrated by homology-directed repair into the genome in frame with the coding region of the gene of interest, resulting in expression of a fusion protein at physiological levels.
This linker is shared by HS and CS/DS chains and its sequence is integrated by xylose-galactose-galactose-GlcA [ 25, 26].
Subsequently, L. johnsonii FI9785 was transformed with pFI2652, and the nisRK sequences were integrated into the chromosome by gene replacement (Maguin et al. 1996) using 30 °C as the permissive temperature and 42 °C as the non-permissive temperature, to create the nisRK-carrying strain FI10744.
In this study, over 1,737 Solexa sequences were integrated in the genetic map through linkage analysis by statistical significance test.
Finally, the 57,307 sequences were integrated into 43,236 unique sequences, i.e. the 43k EST assembly, for use in gene space enrichment by oligonucleotide microarray.
However molecular characterization of these recombinant organisms established that the entire plasmid carrying the transposon sequences was integrated into the A. marginale chromosome by a single crossover homologous recombination mechanism instead of the classical cut and paste mechanism of transposition [ 41].
The results from the promoter sequence enrichment analysis and pathway enrichment analysis were integrated by using protein-protein interaction data with the IPA interaction database.
Yet more variables, such as promotor sequences and protein levels could be integrated by CA, either as done so far, i. e. by applicative arrangement in a two-way table, or by multiple or joint correspondence analysis.
A geneticine resistant and 5-FOA sensitive clone was then transformed with plasmid pToK495 (Ycplac111/pGAL-NOP14), it was selected for resulting clones resistant to 5-FOA on galactose containing medium and a TAP-tag coding sequence was integrated at the NOC4 gene locus by homologous recombination according to [39].
Southern blotting analysis indicated that both lines contained a single GFP insertion, while PCR and DNA sequencing confirmed that the targeting vector had been integrated by homologous recombination (Fig. 1a c).
