title: Construction and evaluation of shuttle vector, pGYC4α, based on pYC2 from Lactobacillus sakei.
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.
authors.affiliation: Department of Food and Nutrition, Chosun University, 375 Seosuk-Dong, Dong-Gu, Gwangju, 501-759, Republic of Korea.
authors.forename: Eun Ju
authors.initials: EJ
authors.lastname: Yang
authors.forename: Hae Choon
authors.initials: HC
authors.lastname: Chang
mesh_terms: D001407:Bacillus / Q000201:enzymology / Q000235:genetics; D005516:Food Microbiology; D005822:Genetic Vectors / Q000235:genetics*; D007778:Lactobacillus / Q000235:genetics*; D008957:Models, Genetic; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D000516:alpha-Amylases / Q000235:genetics / Q000378:metabolism*
chemical_list: D000516:alpha-Amylases