title: [Construction of genomic library of L. interrogans serovar lai using lambda gt11 as the vector and a study of recombiant plasmid pDL121].
abstract: A genomic library of L. interrogans serovar lai strain 017 has been constructed using lambda gt11 as the vector. DNA was partially digested by two blunt-end restriction enzymes, then methylated with EcoR I methylase; after EcoR I linker was added to the DNA, the linker-ended DNA was ligated to the dephosphorylated EcoR I digested lambda gt11 arms. The recombined DNA was packaged in vitro, and used to transduct E. coli Y1090 for amplification. There were 2.1 x 10(6) recombinant bacteriophages as recognized by their ability to form white plaques plated on Lac host in the presence of both IPTG and X-Ga1. A positive clone, designated lambda DL12, was screened with a rabbit anti-serum against L. interrogans serovar lai from the genomic library. The DNA from lambda DL12 was subcloned into plasmid pUC18. A recombinant (designated as pDL121) was obtained. SDS-PAGE analysis indicated that a 23 kd was expressed in E. coli JM 103 harboring pDL121. Western blotting analysis showed that a specific protein band molecular weight of 23 kd could be recognized by the rabbit antiserum against L. interrogans serovar lai strain 017.
authors.affiliation: Research Group of West China University Medical Science, Chengdu.
authors.forename: H
authors.initials: H
authors.lastname: Liu
authors.forename: B
authors.initials: B
authors.lastname: Dai
authors.forename: B
authors.initials: B
authors.lastname: Jing
authors.forename: W
authors.initials: W
authors.lastname: Wu
authors.forename: S
authors.initials: S
authors.lastname: Li
authors.forename: Z
authors.initials: Z
authors.lastname: Fang
authors.forename: H
authors.initials: H
authors.lastname: Zhao
authors.forename: D
authors.initials: D
authors.lastname: Ye
authors.forename: R
authors.initials: R
authors.lastname: Yan
authors.forename: J
authors.initials: J
authors.lastname: Liu
authors.forename: S
authors.initials: S
authors.lastname: Song
authors.forename: Y
authors.initials: Y
authors.lastname: Yang
authors.forename: Y
authors.initials: Y
authors.lastname: Zhang
authors.forename: F
authors.initials: F
authors.lastname: Liu
authors.forename: Y
authors.initials: Y
authors.lastname: Tu
authors.forename: H
authors.initials: H
authors.lastname: Yang
authors.forename: Z
authors.initials: Z
authors.lastname: Huang
authors.forename: L
authors.initials: L
authors.lastname: Liang
authors.forename: L
authors.initials: L
authors.lastname: Hu
authors.forename: M
authors.initials: M
authors.lastname: Zhao
mesh_terms: D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics*; D004274:DNA, Recombinant / Q000096:biosynthesis*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors; D015698:Genomic Library; D007921:Leptospira interrogans / Q000235:genetics*; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D014162:Transfection
chemical_list: D004269:DNA, Bacterial; D004274:DNA, Recombinant; D011993:Recombinant Fusion Proteins