RRC ID 39580
Author Omasa T, Cao Y, Park JY, Takagi Y, Kimura S, Yano H, Honda K, Asakawa S, Shimizu N, Ohtake H.
Title Bacterial artificial chromosome library for genome-wide analysis of Chinese hamster ovary cells.
Journal Biotechnol Bioeng
Abstract Chinese hamster ovary (CHO) cell lines are widely used for scientific research and biotechnology. A CHO genomic bacterial artificial chromosome (BAC) library was constructed from a mouse dihydrofolate reductase (DHFR) gene-amplified CHO DR1000L-4N cell line for genome-wide analysis of CHO cell lines. The CHO BAC library consisted of 122,281 clones and was expected to cover the entire CHO genome five times. A CHO chromosomal map was constructed by fluorescence in situ hybridization (FISH) imaging using BAC clones as hybridization probes (BAC-FISH). Thirteen BAC-FISH marker clones were necessary to identify all the 20 individual chromosomes in a DHFR-deficient CHO DG44 cell line because of the aneuploidy of the cell line. To determine the genomic structure of the exogenous Dhfr amplicon, a 165-kb DNA region containing exogenous Dhfr was cloned from the BAC library using high-density replica (HDR) filters and Southern blot analysis. The nucleotide sequence analysis revealed a novel genomic structure in which the vector sequence containing Dhfr was sandwiched by long inverted sequences of the CHO genome.
Volume 104(5)
Pages 986-94
Published 2009-12-1
DOI 10.1002/bit.22463
PMID 19575438
MeSH Animals CHO Cells* Chromosome Mapping Chromosomes, Artificial, Bacterial* Cricetinae Cricetulus Gene Library* Genetic Vectors Genome* In Situ Hybridization, Fluorescence Mice Recombinant Proteins / genetics Tetrahydrofolate Dehydrogenase / genetics
IF 4.26
Times Cited 23
WOS Category BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Resource
DNA material Chinese hamster genomic BAC library clone (RDB13444)