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PMID: 6209131 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A gene sequence expressed only in undifferentiated EC, EK cells and testes.

The EMBO journal ·Vol. 3 ·No. 10 ·1984-10-00 ·Pages 2279-85

Stacey AJ, Evans MJ

Abstract

A clone, EC1, has been isolated from a cDNA library prepared from 4-day embryoid bodies formed by suspension culture of PSMB EC cells. This clone has been used to screen a variety of RNA sources including adult tissues, embryonal carcinoma (EC), and endoderm cell lines. A 3-kb poly(A)+ RNA species was found to be present only in undifferentiated EC cells and adult mouse testes. This species was significantly reduced in testes of W/Wv mice compared with wild-type at this locus. Germ cells and their progeny are therefore implicated as the source of the RNA in testes. Hybrid-selected RNA from PSMB could be translated in vitro into a 35-kd protein, but no translatable message was evident in either PYS-2 (parietal), or PSA5-E (visceral) endoderm cell lines. DNA sequencing of the EC1 insert revealed that it is 744 bp in length, the 3' 460 bp of which are in open reading frame. Comparison with known sequences have shown no significant homology. EC1 subclones in M13 have been used to generate single-stranded probes for hybridisation to RNA in situ in tissue sections. Hybridisation of the strand complementary to RNA produces a signal limited to the central regions of embryoid bodies formed on suspension culture of embryo-derived EK cells coinciding with the presence of undifferentiated cells. Probing of a mouse genomic library and Southern blots of liver DNA with EC1 reveals that the gene is present as a single copy.

MeSH Terms
Amino Acid Sequence Animals Base Sequence Cell Differentiation DNA/analysis Male Mice Mice, Inbred BALB C Nucleic Acid Hybridization Poly A/metabolism Protein Biosynthesis RNA/analysis,metabolism RNA, Messenger Teratoma/genetics Testis/analysis
Chemicals
RNA, Messenger Poly A RNA DNA
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Stacey A J
Evans M J
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37 references, click to expand
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1984-10-00
Pages
2279-85
Language
English
Region
England
NLM ID
8208664
PMCID
PMC557680
Subset
IM
Databases
GENBANK
X00951
External Links
PubMed source PMC Full Text View journal details
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