The shuttle vector pGYC4 alpha (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, alpha-amylase gene from Bacillus licheniformis containing its own signal peptide, and lactococcal promoter P32. Transformation and expression of a selection marker gene (alpha-amylase) with pGYC4 alpha were demonstrated in Escherichia coli and several lactic acid bacteria (LAB). The highest alpha-amylase activity in LAB transformants was obtained in M17/0.25% glucose media with 0.5% CaCO3. 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.