PMID- 20347775
OWN - NLM
STAT- MEDLINE
DCOM- 20100630
LR  - 20100329
IS  - 1347-4421 (Electronic)
IS  - 1347-4421 (Linking)
VI  - 109
IP  - 5
DP  - 2010 May
TI  - Identification and analysis of specific chromosomal region adjacent to exogenous 
      Dhfr-amplified region in Chinese hamster ovary cell genome.
PG  - 504-11
LID - 10.1016/j.jbiosc.2009.10.019 [doi]
AB  - Chinese hamster ovary (CHO) cells are widely used for the stable production of 
      recombinant proteins. Gene amplification techniques are frequently used to 
      improve of protein production, and the dihydrofolate reductase (DHFR) gene 
      amplification system is most widely used in the CHO cell line. We previously 
      constructed a CHO genomic bacterial artificial chromosome (BAC) library from a 
      mouse Dhfr-amplified CHO DR1000L-4N cell line and one BAC clone (Cg0031N14) 
      containing the CHO genomic DNA sequence adjacent to Dhfr was selected. To 
      identify the specific chromosomal region adjacent to the exogenous Dhfr-amplified 
      region in the CHO cell genome, we performed further screening of BAC clones to 
      obtain other Dhfr-amplified regions in the CHO genome. From the screening by 
      high-density replica filter hybridization using a digoxigenin-labeled 
      pSV2-dhfr/hGM-CSF probe, we obtained 8 new BAC clones containing a Dhfr-amplified 
      region. To define the structures of the 8 BAC clones, Southern blot analysis, BAC 
      end sequencing and fluorescence in situ hybridization (FISH) were performed. 
      These results revealed that all the selected BAC clones contained a large 
      palindrome structure with a small inverted repeat in the junction region. This 
      suggests that the obtained amplicon structure in the Dhfr-amplified region in the 
      CHO genome plays an important role in exogenous gene amplification.
CI  - (c) 2009 The Society for Biotechnology, Japan. Published by Elsevier B.V. All 
      rights reserved.
FAU - Park, Joon Young
AU  - Park JY
AD  - Department of Biotechnology, Graduate School of Engineering, Osaka University, 
      2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
FAU - Takagi, Yasuhiro
AU  - Takagi Y
FAU - Yamatani, Miyuki
AU  - Yamatani M
FAU - Honda, Kohsuke
AU  - Honda K
FAU - Asakawa, Shuichi
AU  - Asakawa S
FAU - Shimizu, Nobuyoshi
AU  - Shimizu N
FAU - Omasa, Takeshi
AU  - Omasa T
FAU - Ohtake, Hisao
AU  - Ohtake H
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20091118
PL  - Japan
TA  - J Biosci Bioeng
JT  - Journal of bioscience and bioengineering
JID - 100888800
RN  - EC 1.5.1.3 (Tetrahydrofolate Dehydrogenase)
SB  - IM
MH  - Animals
MH  - Base Sequence
MH  - CHO Cells/*physiology
MH  - *Chromosome Mapping
MH  - Cricetinae/*genetics
MH  - Cricetulus/*genetics
MH  - Gene Amplification/*genetics
MH  - Genome/*genetics
MH  - Molecular Sequence Data
MH  - Tetrahydrofolate Dehydrogenase/*genetics
EDAT- 2010/03/30 06:00
MHDA- 2010/07/01 06:00
CRDT- 2010/03/30 06:00
PHST- 2009/10/02 00:00 [received]
PHST- 2009/10/19 00:00 [revised]
PHST- 2009/10/24 00:00 [accepted]
PHST- 2010/03/30 06:00 [entrez]
PHST- 2010/03/30 06:00 [pubmed]
PHST- 2010/07/01 06:00 [medline]
AID - S1389-1723(09)01002-0 [pii]
AID - 10.1016/j.jbiosc.2009.10.019 [doi]
PST - ppublish
SO  - J Biosci Bioeng. 2010 May;109(5):504-11. doi: 10.1016/j.jbiosc.2009.10.019. Epub 
      2009 Nov 18.