| 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 |
4.002
|
| Times Cited |
23
|
|
WOS Category
|
BIOTECHNOLOGY & APPLIED MICROBIOLOGY
|
| Resource |
| DNA material |
Chinese hamster genomic BAC library clone (RDB13444) |
