Novel genes expressed in the chick otocyst during development: identification using differential display of RNA.
Differential display of mRNA is a technique that enables the researcher to compare genes expressed in two or more different tissues or in the same tissue or cell under different conditions. The method is based on polymerase chain reaction amplification and comparison of specific subsets of mRNA. We have used this method to clone partial complementary DNAs (cDNAs; amplicons) for genes expressed in the otocyst in order to identify genes that may be involved in development of the inner ear. A full length cDNA was isolated from an embryonic quail head library with an amplicon (KH121) obtained from the otocyst. This avian cDNA encoded a novel, 172-amino acid acidic protein and detected a major transcript of ca 0.8 kb in RNA from chick embryos and several neonatal chick tissues. The full length avian cDNA had high sequence identity to several human cDNAs (expressed sequence tags) from human fetal tissues, including cochlea, brain, liver/spleen and lung, and from placenta. The human homologue of the avian gene encoded a protein that was 183 amino acids long and had 75.6% amino acid sequence identity to the avian protein. These results identified both the avian and human homologues of an evolutionarily conserved gene encoding a small acidic protein of unknown function; however, expression of this gene was not restricted to otocysts.
Gong
T W
TW
Kresge Hearing Research Institute, Department of Otolaryngology/Head-Neck Surgery, University of Michigan Medical School, Ann Arbor 48109-0648, USA.
Hegeman
A D
AD
Shin
J J
JJ
Lindberg
K H
KH
Barald
K F
KF
Lomax
M I
MI
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D002642:Chick Embryo; D003051:Cochlea / Q000166:cytology / Q000254:growth & development* / Q000502:physiology; D017124:Conserved Sequence; D017931:DNA Primers; D015342:DNA Probes; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000096:biosynthesis; D016133:Polymerase Chain Reaction; D011784:Quail; D012333:RNA, Messenger / Q000096:biosynthesis* / Q000302:isolation & purification; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D015342:DNA Probes; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger