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  • Title: Construction and evaluation of shuttle vector, pGYC4α, based on pYC2 from Lactobacillus sakei.
    Author: Yang EJ, Chang HC.
    Journal: Biotechnol Lett; 2011 Mar; 33(3):599-605. PubMed ID: 21072560.
    Abstract:
    The shuttle vector pGYC4α (6,157 bp) was constructed based on the sigma-replicon plasmid pYC2 from Lactobacillus sakei BM5 isolated from kimchi. The vector contained inserts of the ColE1 replicon, α-amylase gene from Bacillus licheniformis containing its own signal peptide, and lactococcal promoter P32. Transformation and expression of a selection marker gene (α-amylase) with pGYC4α were demonstrated in Escherichia coli and several lactic acid bacteria (LAB). The highest α-amylase activity in LAB transformants was obtained in M17/0.25% glucose media with 0.5% CaCO(3). The segregational stability of the shuttle vector in LAB was 100% for more than 100 generations in the absence of antibiotic pressure. The developed vector might be useful as a genetic tool for food industries.
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