PMID- 10402672
OWN - NLM
STAT- MEDLINE
DCOM- 19990812
LR  - 20151119
IS  - 0192-253X (Print)
IS  - 0192-253X (Linking)
VI  - 25
IP  - 1
DP  - 1999
TI  - Gene trap screening using negative selection: identification of two tandem,
      differentially expressed loci with potential hematopoietic function.
PG  - 49-63
AB  - A fusion gene between Escherichia coli lacZ and herpes simplex virus thymidine
      kinase (HSV-tk) was constructed and used in a gene trap screen for hematopoietic 
      loci in mouse embryonic stem (ES) cells. This gene, galtek, allowed both
      convenient histochemical detection of expression as well as ablation of
      expressing cells under 1-(2-deoxy-2-fluoro-beta-D-arabinofuranosyl)-5-iodouracil 
      (FIAU) selection. Individual ES cell clones bearing gene trap insertions were
      differentiated in the presence of FIAU and scored for erythropoietic activity at 
      day 9 of differentiation. Screening of a total of 235 independent gene trap lines
      identified one clone, F3, which consistently demonstrated FIAU-sensitive
      erythropoiesis during in vitro differentiation. Cloning of endogenous transcribed
      sequences from the F3 insertion site identified two distinct transcription units,
      F3-1 and F3-2, encoding mRNAs of approximately 1.3 kb and 3.35 kb, respectively. 
      The transcripts were unrelated and did not exhibit similarity to known sequences.
      Both loci demonstrated similar relative levels of expression in the heart,
      testis, kidney, and lung as assessed by Northern blot hybridization. Whole-mount 
      in situ hybridization detected F3-2 expression at multiple sites in embryonic day
      (E) 10.5 embryos, including the genital ridges, the aortic endothelium, and
      endothelium-associated cell clusters within the aortic lumen. Expression of F3-2 
      in the aortic endothelium and endothelium-associated clusters overlapped that of 
      gata-2, a gene required for hematopoietic development. The FIAU sensitivity of
      hematopoiesis in F3 embryoid bodies may result from expression of galtek during
      the formation of early hematopoietic cells, directed by regulatory signals from
      one or both of these endogenous loci.
FAU - Cannon, J P
AU  - Cannon JP
AD  - Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, 
      Texas 77030, USA.
FAU - Colicos, S M
AU  - Colicos SM
FAU - Belmont, J W
AU  - Belmont JW
LA  - eng
GR  - HD36280/HD/NICHD NIH HHS/United States
GR  - HL51232/HL/NHLBI NIH HHS/United States
GR  - T-32-AI07495-03/05/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Dev Genet
JT  - Developmental genetics
JID - 7909963
RN  - 0 (Biomarkers)
RN  - 0 (DNA, Complementary)
RN  - 0 (DNA-Binding Proteins)
RN  - 0 (GATA2 Transcription Factor)
RN  - 0 (Gata2 protein, mouse)
RN  - 0 (Gtlf3a protein, mouse)
RN  - 0 (Gtlf3b protein, mouse)
RN  - 0 (Proteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (Transcription Factors)
RN  - EC 2.7.1.21 (Thymidine Kinase)
RN  - EC 3.2.1.23 (beta-Galactosidase)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - *Biomarkers
MH  - DNA, Complementary
MH  - DNA-Binding Proteins/genetics
MH  - GATA2 Transcription Factor
MH  - Gene Expression Regulation, Developmental
MH  - Hematopoiesis/*genetics
MH  - Mice
MH  - Mice, Inbred C57BL
MH  - Mice, Inbred CBA
MH  - Molecular Sequence Data
MH  - Polymerase Chain Reaction
MH  - Proteins/*genetics
MH  - RNA, Messenger/genetics
MH  - *Selection, Genetic
MH  - Thymidine Kinase/genetics
MH  - Transcription Factors/genetics
MH  - beta-Galactosidase/genetics
EDAT- 1999/07/14 10:00
MHDA- 2000/06/20 09:00
CRDT- 1999/07/14 10:00
PHST- 1999/07/14 10:00 [pubmed]
PHST- 2000/06/20 09:00 [medline]
PHST- 1999/07/14 10:00 [entrez]
AID - 10.1002/(SICI)1520-6408(1999)25:1<49::AID-DVG6>3.0.CO;2-S [pii]
AID - 10.1002/(SICI)1520-6408(1999)25:1<49::AID-DVG6>3.0.CO;2-S [doi]
PST - ppublish
SO  - Dev Genet. 1999;25(1):49-63. doi:
      10.1002/(SICI)1520-6408(1999)25:1<49::AID-DVG6>3.0.CO;2-S.