PMID- 10037815
OWN - NLM
STAT- MEDLINE
DCOM- 19990427
LR  - 20190501
IS  - 0305-1048 (Print)
IS  - 0305-1048 (Linking)
VI  - 27
IP  - 6
DP  - 1999 Mar 15
TI  - Combining SSH and cDNA microarrays for rapid identification of differentially
      expressed genes.
PG  - 1517-23
AB  - Comparing patterns of gene expression in cell lines and tissues has important
      applications in a variety of biological systems. In this study we have examined
      whether the emerging technology of cDNA microarrays will allow a high throughput 
      analysis of expression of cDNA clones generated by suppression subtractive
      hybridization (SSH). A set of cDNA clones including 332 SSH inserts amplified by 
      PCR was arrayed using robotic printing. The cDNA arrays were hybridized with
      fluorescent labeled probes prepared from RNA from ER-positive (MCF7 and T47D) and
      ER-negative (MDA-MB-231 and HBL-100) breast cancer cell lines. Ten clones were
      identified that were over-expressed by at least a factor of five in the
      ER-positive cell lines. Northern blot analysis confirmed over-expression of these
      10 cDNAs. Sequence analysis identified four of these clones as cytokeratin 19,
      GATA-3, CD24 and glutathione-S-transferase mu-3. Of the remaining six cDNA
      clones, four clones matched EST sequences from two different genes and two clones
      were novel sequences. Flow cytometry and immunofluorescence confirmed that CD24
      protein was over-expressed in the ER-positive cell lines. We conclude that SSH
      and microarray technology can be successfully applied to identify differentially 
      expressed genes. This approach allowed the identification of differentially
      expressed genes without the need to obtain previously cloned cDNAs.
FAU - Yang, G P
AU  - Yang GP
AD  - Department of Surgery and Department of Biochemistry, Stanford University,
      Stanford, CA 94305-5414, USA.
FAU - Ross, D T
AU  - Ross DT
FAU - Kuang, W W
AU  - Kuang WW
FAU - Brown, P O
AU  - Brown PO
FAU - Weigel, R J
AU  - Weigel RJ
LA  - eng
PT  - Journal Article
PL  - England
TA  - Nucleic Acids Res
JT  - Nucleic acids research
JID - 0411011
RN  - 0 (Antigens, CD)
RN  - 0 (CD24 Antigen)
RN  - 0 (CD24 protein, human)
RN  - 0 (DNA, Complementary)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (GATA3 Transcription Factor)
RN  - 0 (GATA3 protein, human)
RN  - 0 (Membrane Glycoproteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (Receptors, Estrogen)
RN  - 0 (Trans-Activators)
RN  - 68238-35-7 (Keratins)
RN  - EC 2.5.1.18 (Glutathione Transferase)
SB  - IM
MH  - Antigens, CD/genetics/isolation & purification
MH  - Blotting, Northern
MH  - Breast Neoplasms/*genetics
MH  - CD24 Antigen
MH  - Cloning, Molecular
MH  - *DNA, Complementary
MH  - DNA-Binding Proteins/genetics
MH  - Expressed Sequence Tags
MH  - Female
MH  - GATA3 Transcription Factor
MH  - Gene Expression
MH  - Gene Library
MH  - Glutathione Transferase/genetics
MH  - Humans
MH  - Keratins/genetics
MH  - *Membrane Glycoproteins
MH  - Nucleic Acid Hybridization/*methods
MH  - Oligonucleotide Array Sequence Analysis/*methods
MH  - RNA, Messenger/*isolation & purification
MH  - Receptors, Estrogen/isolation & purification
MH  - Sequence Analysis, DNA
MH  - Trans-Activators/genetics
MH  - Tumor Cells, Cultured
PMC - PMC148347
EDAT- 1999/02/26 00:00
MHDA- 1999/02/26 00:01
CRDT- 1999/02/26 00:00
PHST- 1999/02/26 00:00 [pubmed]
PHST- 1999/02/26 00:01 [medline]
PHST- 1999/02/26 00:00 [entrez]
AID - gkc278 [pii]
AID - 10.1093/nar/27.6.1517 [doi]
PST - ppublish
SO  - Nucleic Acids Res. 1999 Mar 15;27(6):1517-23. doi: 10.1093/nar/27.6.1517.