PMID- 7568467
OWN - NLM
STAT- MEDLINE
DCOM- 19951023
LR  - 20131121
IS  - 0147-619X (Print)
IS  - 0147-619X (Linking)
VI  - 33
IP  - 3
DP  - 1995 May
TI  - An episomal expression vector system for monitoring sequence-specific effects on 
      mRNA stability in human cell lines.
PG  - 198-207
AB  - A plasmid expression system has been developed which allows sequence-specific 
      effects on mRNA degradation rates to be determined. This system uses stable, 
      nonintegrating vectors that provide consistent levels of mRNA expression without 
      the position effects common to integrating vectors. cDNAs encoding putative 
      instability elements may be subcloned into the 5' untranslated region (5'UTR), 
      the coding region, or the proximal 3'UTR of a beta-globin cDNA reporter. The 
      effects of these sequences on mRNA stability may then be determined by 
      actinomycin time course analyses of the fusion mRNAs and recombinant beta-globin 
      mRNA in human cell lines. To demonstrate the utility of the vector system we 
      fused an 820-bp fragment of the cDNA encoding the proximal 3'UTR of human 
      3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase to the 3'UTR of the 
      beta-globin reporter and introduced the vector into the human hepatocarcinoma 
      cell line, HepG2. The fusion mRNA was degraded at a rate 2- to 2.5-fold greater 
      than that of beta-globin alone, at a rate similar to that reported for HMG CoA 
      reductase mRNA in normal rat liver. Similar to a number of other relatively 
      unstable mRNAs, the rate of fusion mRNA degradation was greatly decreased by 
      treatment with cycloheximide.
FAU - Wilson, G M
AU  - Wilson GM
AD  - Department of Biochemistry and Cancer Research Laboratories, Queen's University, 
      Kingston, Ontario, Canada.
FAU - Deeley, R G
AU  - Deeley RG
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Plasmid
JT  - Plasmid
JID - 7802221
RN  - 0 (DNA, Complementary)
RN  - 0 (RNA, Messenger)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 1CC1JFE158 (Dactinomycin)
RN  - 63231-63-0 (RNA)
RN  - 9004-22-2 (Globins)
RN  - EC 1.1.1.- (Hydroxymethylglutaryl CoA Reductases)
SB  - IM
MH  - Base Sequence
MH  - Blotting, Northern
MH  - Carcinoma, Hepatocellular
MH  - Cell Line
MH  - Cloning, Molecular
MH  - DNA, Complementary
MH  - Dactinomycin/pharmacology
MH  - Gene Expression/drug effects
MH  - *Genetic Vectors
MH  - Globins/*biosynthesis
MH  - Humans
MH  - Hydroxymethylglutaryl CoA Reductases/*genetics
MH  - Liver Neoplasms
MH  - Molecular Sequence Data
MH  - *Plasmids
MH  - RNA/isolation & purification
MH  - RNA, Messenger/biosynthesis/isolation & purification/*metabolism
MH  - Recombinant Fusion Proteins/biosynthesis
MH  - Recombinant Proteins/biosynthesis
MH  - Repetitive Sequences, Nucleic Acid
MH  - Restriction Mapping
MH  - Transfection/*methods
MH  - Tumor Cells, Cultured
EDAT- 1995/05/01 00:00
MHDA- 1995/05/01 00:01
CRDT- 1995/05/01 00:00
PHST- 1995/05/01 00:00 [pubmed]
PHST- 1995/05/01 00:01 [medline]
PHST- 1995/05/01 00:00 [entrez]
AID - S0147-619X(85)71021-9 [pii]
AID - 10.1006/plas.1995.1021 [doi]
PST - ppublish
SO  - Plasmid. 1995 May;33(3):198-207. doi: 10.1006/plas.1995.1021.