AI Article Synopsis
- The vector pIRK was created using a fragment of rDNA from Kluyveromyces lactis and included the URA3 gene from Saccharomyces cerevisiae for selection.
- In K. lactis transformants, pIRK had an average of 120 copies per cell and demonstrated high stability after 50 generations, with all integrations found on chromosome IV.
- The LAC4 gene from K. fragilis was expressed using pIRK, resulting in beta-galactosidase production that was 8.6 times greater than the wild type strain under the same conditions.
Article Abstract
The vector pIRK was constructed by using a 2.2 kb EcoRI fragment of rDNA from Kluyveromyces lactis for targeted homologous recombination, with the URA3 gene from Saccharomyces cerevisiae acting as a selection marker. By the examination of the copy number, stability and chromosomal location of the vector in K. lactis transformants the results demonstrated that: (1) of the different transformants, the average copy number of the plasmid pIRK was 120 per cell; (2) after 50 generations of growth in rich medium, the vector displayed high stability. (3) all integration events occurred in the chromosome IV where genomic rDNA located. Using this vector, the LAC4 gene cloned from K. fragilis was expressed. The yield of beta-galactosidase related directly to the vector's copy number. The highest activity of beta-galactosidase produced by transformants was 8.6 times higher than that produced by the wild type strain of K. fragilis under the same conditions.
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