title: An arrayed human genomic library constructed in the PAC shuttle vector pJCPAC-Mam2 for genome-wide association studies and gene therapy.
abstract: The various iterations of the HapMap Project and many genome-wide association studies (GWAS) have identified hundreds of potential genes involved in monogenic and multifactorial traits. We constructed an arrayed 115,000-member human genomic library in the PAC shuttle vector pJCPAC-Mam2 that can be propagated in both bacterial and human cells. The library appears to represent a two-fold coverage of the human genome. Transient transfection of a p53-containing PAC clone into p53-null Saos-2 human osteosarcoma cells demonstrated that both p53 mRNA and protein were produced. Additionally, expression of the p53 protein triggered apoptosis in a subset of the Saos-2 cells. This library should serve as a valuable resource to validate potential disease genes identified by GWAS in human cell lines and in animal models. Also, individual library members could potentially be used for gene therapy trials for a variety of recessive disorders.
authors.affiliation: Department of Biology, Elizabethtown College, Elizabethtown, PA 17022, USA.
authors.forename: John
authors.initials: J
authors.lastname: Fuesler
authors.forename: Yasunori
authors.initials: Y
authors.lastname: Nagahama
authors.forename: Joseph
authors.initials: J
authors.lastname: Szulewski
authors.forename: Joshua
authors.initials: J
authors.lastname: Mundorff
authors.forename: Stephanie
authors.initials: S
authors.lastname: Bireley
authors.forename: Jonathon S
authors.initials: JS
authors.lastname: Coren
mesh_terms: D017209:Apoptosis; D045744:Cell Line, Tumor; D003001:Cloning, Molecular; D016158:Genes, p53; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D055106:Genome-Wide Association Study / Q000379:methods*; D015698:Genomic Library; D023281:Genomics / Q000379:methods; D049452:Green Fluorescent Proteins / Q000737:chemistry; D060148:HapMap Project; D006801:Humans; D012516:Osteosarcoma / Q000378:metabolism; D014176:Protein Biosynthesis; D014158:Transcription, Genetic; D016159:Tumor Suppressor Protein p53 / Q000235:genetics
chemical_list: C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; D049452:Green Fluorescent Proteins