PMID- 10592510
OWN - NLM
STAT- MEDLINE
DCOM- 20000216
LR  - 20181130
IS  - 0006-3592 (Print)
IS  - 0006-3592 (Linking)
VI  - 67
IP  - 2
DP  - 2000 Jan 20
TI  - Engineering CHO cells to overexpress a secreted reporter protein upon induction 
      from mouse mammary tumor virus promoter.
PG  - 134-40
AB  - The mouse mammary tumor virus (MMTV) promoter is induced by the addition of a 
      glucocorticoid hormone or analog such as dexamethasone. The hormone binds to its 
      specific transcription factor, glucocorticoid receptor (GR), and the activated 
      complex then binds to the glucocorticoid response elements (GREs) in the enhancer 
      region of the MMTV promoter to induce the overexpression of downstream genes. We 
      have constructed an expression vector for a reporter protein, secreted alkaline 
      phosphatase (SEAP),controlled by the MMTV promoter and co-transfected this vector 
      along with a GR expression cassette into Chinese hamster ovary (CHO) cells. High 
      producers were cloned and grown in suspension cultures. A very high titer, over 
      0.4 mg/mL, of SEAP was obtained from this inducible overexpression system, about 
      ten times that achievable with the same reporter protein using the strong 
      constitutive SV40 promoter in CHO cells. A peak production rate of 187 pg SEAP 
      per cell per day was observed within 3 days after induction, compared to the peak 
      rate of 23 pg SEAP per cell per day expressed using the constitutive SV40 
      promoter. With the reduced or zero growth rate during the protein production 
      phase, this novel MMTV overexpression system is highly suited for optimizing 
      glycoprotein synthesis rates in high cell density fed-batch or perfusion 
      bioreactors.
CI  - Copyright 2000 John Wiley & Sons, Inc.
FAU - James, R I
AU  - James RI
AD  - Department of Chemical Engineering, University of Colorado, Boulder, Colorado 
      80309-0424, USA.
FAU - Elton, J P
AU  - Elton JP
FAU - Todd, P
AU  - Todd P
FAU - Kompala, D S
AU  - Kompala DS
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - United States
TA  - Biotechnol Bioeng
JT  - Biotechnology and bioengineering
JID - 7502021
RN  - 0 (Culture Media)
RN  - 0 (Receptors, Glucocorticoid)
RN  - 0 (Recombinant Proteins)
RN  - EC 3.1.3.1 (Alkaline Phosphatase)
SB  - IM
MH  - Alkaline Phosphatase/genetics/metabolism
MH  - Animals
MH  - CHO Cells/*physiology
MH  - Cricetinae
MH  - Culture Media
MH  - *Genes, Reporter
MH  - Genetic Engineering/*methods
MH  - Genetic Vectors
MH  - Mammary Tumor Virus, Mouse/*genetics
MH  - Promoter Regions, Genetic
MH  - Protein Engineering/methods
MH  - Receptors, Glucocorticoid/genetics/metabolism
MH  - Recombinant Proteins/*genetics/*metabolism
MH  - Simian virus 40/genetics
MH  - Transfection
EDAT- 1999/12/11 09:00
MHDA- 2000/02/19 09:00
CRDT- 1999/12/11 09:00
PHST- 1999/12/11 09:00 [pubmed]
PHST- 2000/02/19 09:00 [medline]
PHST- 1999/12/11 09:00 [entrez]
AID - 10.1002/(SICI)1097-0290(20000120)67:2<134::AID-BIT2>3.0.CO;2-V [pii]
PST - ppublish
SO  - Biotechnol Bioeng. 2000 Jan 20;67(2):134-40.