Human BAC library: construction and rapid screening

Asakawa, Shuichi; Abe, Izumi; Kudoh, Yoshiki; Kishi, Noriyuki; Wang, Yimin; Kubota, Ryo; Kudoh, Jun; Kawasaki, Kazuhiko; Minoshima, Shinsei; Shimizu, Nobuyoshi

doi:10.1016/s0378-1119(97)00044-9

Cited by 174 publications

(118 citation statements)

References 31 publications

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“…Cloning of the pBD-1 Gene-A two-step polymerase chain reaction (PCR) was used to screen a porcine genomic bacterial artificial chromosome (BAC) library 2 for BAC clones containing the pBD-1 gene as described previously (45). Two pairs of primers derived from the cDNA or partial genomic sequence of pBD-1 were used: 5Ј-ATG AGA CTC CAC CGC CTC CTC CT-3Ј (sense) and 5Ј-GCA GCA TTT GAC TTG GGG CAT G-3Ј (antisense) (product size: 1.7 kb); and 5Ј-GGG TTA TGA AGT ATC CAT GTA ACC GAC-3Ј (sense) and 5Ј-GGT CAG CTC CAC TAG GAC AGC AGA G-3Ј (antisense) (product size: 812 bp).…”

Section: Methodsmentioning

confidence: 99%

Cloning and Characterization of the Gene for a New Epithelial β-Defensin

Zhang

Hiraiwa

Yasue

et al. 1999

Journal of Biological Chemistry

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Mammalian ␤-defensins are endogenous cysteine-rich peptide antibiotics that are produced either by epithelial cells lining the respiratory, digestive, and urogenital tracts or by granulocytes and macrophages. A growing body of evidence has implicated these peptides in host defense, particularly mucosal innate immunity. We previously reported the cloning of the full-length cDNA for a porcine ␤-defensin (pBD-1), which was found to be expressed throughout the airway and oral mucosa. Here, we provide the structural organization of the pBD-1 gene, showing that the entire gene spans ϳ1.9 kilobases with two short exons separated by a 1.5-kilobase intron. Fluorescence in situ hybridization mapped the pBD-1 gene to porcine chromosome 15q14-q15.1 within a region of conserved synteny to the chromosomal locations of human and mouse ␣-and ␤-defensins. We also provide several independent lines of evidence showing that the pBD-1 gene is expressed constitutively during inflammation and infection, despite its resemblance to many inducible epithelial ␤-defensins in amino acid sequence, genomic structure, and sites of expression. First, stimulation of primary porcine tongue epithelial cells with lipopolysaccharide, tumor necrosis factor-␣, and interleukin (IL)-1␤ failed to up-regulate the expression of pBD-1 mRNA. Second, pBD-1 gene expression was not enhanced in either digestive or respiratory mucosa of pigs following a 2-day infection with Salmonella typhimurium or Actinobacillus pleuropneumoniae. Last, direct transfection of the pBD-1 gene promoter into NIH/3T3 cells showed no difference in reporter gene activity in response to stimulation by lipopolysaccharide and IL-1␤. The constitutive expression of pBD-1 in airway and oral mucosa, which is consistent with a lack of consensus binding sites for nuclear factor-B or NF-IL-6 in its promoter region, suggests that it may play a surveillance role in maintaining the steady state of microflora on mucosal surfaces.

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Section: Methodsmentioning

confidence: 99%

Cloning and Characterization of the Gene for a New Epithelial β-Defensin

Zhang

Hiraiwa

Yasue

et al. 1999

Journal of Biological Chemistry

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“…This clone was used for FISH analysis (see below). BAC clones were obtained by screening two total human BAC libraries; one was constructed by Asakawa et al (1997), and the other was purchased from Research Genetics (Huntsville, AL, USA). From the latter, we prepared a PCR-based BAC screening system and isolated BAC clones.…”

Section: Isolation Of the Genomic Clones Harbouring The Commonly Delementioning

confidence: 99%

Detailed deletion mapping of chromosome band 14q32 in human neuroblastoma defines a 1.1-Mb region of common allelic loss

et al. 2000

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Neuroblastoma (NB) is a well-known malignant disease in infants, but its molecular mechanisms have not yet been fully elucidated. To investigate the genetic contribution of abnormalities on the long arm of chromosome 14 (14q) in NB, we analysed loss of heterozygosity (LOH) in 54 primary NB samples using 12 microsatellite markers on 14q32. Seventeen (31%) of 54 tumours showed LOH at one or more of the markers analysed, and the smallest common region of allelic loss was identified between D14S62 and D14S987. This region was estimated to be 1-cM long from the linkage map. Fluorescence in situ hybridization also confirmed the loss. There was no statistical correlation between LOH and any clinicopathologic features, including age, stage, amplification of MYCN and ploidy. We further constructed a contig spanning the lost region using bacterial artificial chromosome and estimated this region to be approximately 1.1-Mb by pulsed-field gel electrophoresis. Our results will contribute to cloning and characterizing the putative tumour-associated gene(s) in 14q32 in NB. © 2000 Cancer Research Campaign

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“…We thought that this microsatellite marker might be located within the disease gene. By screening the Keio BAC library (Asakawa et al 1997) using D6S305, we cloned a cDNA consisting of 2960 base pairs, of which 1395 base pairs constituted the open reading frame (Kitada et al 1998). As this was a novel gene, we named it parkin.…”

Section: Progress In Familial Pdmentioning

confidence: 99%

Progress in the pathogenesis and genetics of Parkinson's disease

Mizuno

Hattori

Kubo

et al. 2008

Phil. Trans. R. Soc. B

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Recent progresses in the pathogenesis of sporadic Parkinson's disease (PD) and genetics of familial PD are reviewed. There are common molecular events between sporadic and familial PD, particularly between sporadic PD and PARK1-linked PD due to a-synuclein (SNCA) mutations. In sporadic form, interaction of genetic predisposition and environmental factors is probably a primary event inducing mitochondrial dysfunction and oxidative damage resulting in oligomer and aggregate formations of a-synuclein. In PARK1-linked PD, mutant a-synuclein proteins initiate the disease process as they have increased tendency for self-aggregation. As highly phosphorylated aggregated proteins are deposited in nigral neurons in PD, dysfunctions of proteolytic systems, i.e. the ubiquitin-proteasome system and autophagy-lysosomal pathway, seem to be contributing to the final neurodegenerative process. Studies on the molecular mechanisms of nigral neuronal death in familial forms of PD will contribute further on the understanding of the pathogenesis of sporadic PD.

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Human BAC library: construction and rapid screening

Cited by 174 publications

References 31 publications

Cloning and Characterization of the Gene for a New Epithelial β-Defensin

Cloning and Characterization of the Gene for a New Epithelial β-Defensin

Detailed deletion mapping of chromosome band 14q32 in human neuroblastoma defines a 1.1-Mb region of common allelic loss

Progress in the pathogenesis and genetics of Parkinson's disease

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