title: Construction of a shuttle vector based on the small cryptic plasmid pJY33 from Weissella cibaria 33.
abstract: A cryptic plasmid, pJY33, from Weissella cibaria 33 was characterized. pJY33 was 2365 bp in size with a GC content of 41.27% and contained two putative open reading frames (ORFs). orf1 encoded a putative hypothetical protein of 134 amino acids. orf2 was 849 bp in size, and its putative translation product exhibited 87% identity with a replication initiation factor from a plasmid from W. cibaria KLC140. A Weissella-Escherichia coli shuttle vector, pJY33E (6.5 kb, Em(r)), was constructed by ligation of pJY33 with pBluescript II SK(-) and an erythromycin resistance gene (Em(r)). pJY33E replicated in Lactococcus lactis, Leuconostoc citreum, Lactobacillus brevis, Lactobacillus plantarum, and Weissella confusa. A single-stranded DNA intermediate was detected from Lb. brevis 2.14 harbouring pJY33E, providing evidence for rolling-circle replication of pJY33. Most Lb. brevis 2.14 cells (85.9%) retained pJY33E after one week of daily culturing in MRS broth without Em. An aga gene encoding α-galactosidase (α-Gal) from Leuconostoc mesenteroides was successfully expressed in Lb. brevis 2.14 using pJY33E, and the highest level of α-Gal activity (36.13 U/mg protein) was observed when cells were grown on melibiose.
authors.affiliation: Division of Applied Life Science (BK21 Plus), Graduate School, 501 Jinju-daero, Jinju, Gyeongnam 660-701, Republic of Korea.
authors.forename: Ji Yeong
authors.initials: JY
authors.lastname: Park
authors.affiliation: Division of Applied Life Science (BK21 Plus), Graduate School, 501 Jinju-daero, Jinju, Gyeongnam 660-701, Republic of Korea.
authors.forename: Seon-Ju
authors.initials: SJ
authors.lastname: Jeong
authors.affiliation: Division of Applied Life Science (BK21 Plus), Graduate School, 501 Jinju-daero, Jinju, Gyeongnam 660-701, Republic of Korea.
authors.forename: Hyun Deok
authors.initials: HD
authors.lastname: Sa
authors.affiliation: Division of Applied Life Science (BK21 Plus), Graduate School, 501 Jinju-daero, Jinju, Gyeongnam 660-701, Republic of Korea.
authors.forename: Jae Yong
authors.initials: JY
authors.lastname: Lee
authors.affiliation: Division of Applied Life Science (BK21 Plus), Graduate School, 501 Jinju-daero, Jinju, Gyeongnam 660-701, Republic of Korea.
authors.forename: Xiaoming
authors.initials: X
authors.lastname: Liu
authors.affiliation: Institute of Agriculture and Life Science, Gyeongsang National University, 501 Jinju-daero, Jinju, Gyeongnam 660-701, Republic of Korea.
authors.forename: Min Jeong
authors.initials: MJ
authors.lastname: Cho
authors.affiliation: Institute of Agriculture and Life Science, Gyeongsang National University, 501 Jinju-daero, Jinju, Gyeongnam 660-701, Republic of Korea.
authors.forename: Kang Wook
authors.initials: KW
authors.lastname: Lee
authors.affiliation: Division of Applied Life Science (BK21 Plus), Graduate School, 501 Jinju-daero, Jinju, Gyeongnam 660-701, Republic of Korea; Institute of Agriculture and Life Science, Gyeongsang National University, 501 Jinju-daero, Jinju, Gyeongnam 660-701, Republic of Korea. Electronic address: jeonghkm@gnu.ac.kr.
authors.forename: Jeong Hwan
authors.initials: JH
authors.lastname: Kim
mesh_terms: D001482:Base Composition; D004261:DNA Replication; D004269:DNA, Bacterial / Q000235:genetics; D004917:Erythromycin / Q000494:pharmacology; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D048191:Lactiplantibacillus plantarum / Q000235:genetics; D013294:Lactococcus lactis / Q000235:genetics; D007934:Leuconostoc / Q000235:genetics; D016366:Open Reading Frames; D010957:Plasmids / Q000235:genetics*; D017422:Sequence Analysis, DNA; D058836:Weissella / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D004917:Erythromycin
keywords: Rolling circle replication; Shuttle vector; Weissella cibaria; pJY33E