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. 1987 Mar;84(6):1704–1708. doi: 10.1073/pnas.84.6.1704

Molecular cloning and sequencing of a cDNA for olfactory marker protein.

K E Rogers, P Dasgupta, U Gubler, M Grillo, Y S Khew-Goodall, F L Margolis
PMCID: PMC304505  PMID: 3470751

Abstract

cDNA clones corresponding to mRNA for rat olfactory marker protein (OMP) were isolated from a cDNA library. The library was constructed from olfactory mucosa poly(A)+ RNA enriched for OMP mRNA and cloned into a pBR322-derived plasmid, pMG5. OMP cDNA clones were detected by using a 17-base oligonucleotide probe that contained all 16 possible sequences coding for a known partial amino acid sequence of rat OMP. The identity of these clones was confirmed by hybrid-selected translation and nucleotide sequencing. The sequence of one clone was determined and contained the complete OMP coding region of 486 nucleotides followed by 1630 nucleotides of the 3' untranslated region. The 3' untranslated region included the polyadenylylation signal 16 nucleotides upstream of the poly(A) tail. No other ATG-initiated open reading frame larger than 20 codons was present in register. RNA blot analysis of olfactory mucosa poly(A)+ RNA using this clone as a probe indicated that the level of OMP mRNA, but not its size, declined significantly within a few days following olfactory bulbectomy. OMP mRNA was not detected in 14 nonolfactory rat tissues. Surprisingly, a small amount of OMP mRNA was observed in olfactory bulb. The presence of OMP mRNA in olfactory bulb was confirmed by in vitro translation and immunoprecipitation. These results suggest either that a previously undescribed population of neurons in the olfactory bulb synthesize OMP or that OMP mRNA is transported to the bulb by axonal transport.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barber P. C., Jensen S., Zimmer J. Differentiation of neurons containing olfactory marker protein in adult rat olfactory epithelium transplanted to the anterior chamber of the eye. Neuroscience. 1982;7(11):2687–2695. doi: 10.1016/0306-4522(82)90092-6. [DOI] [PubMed] [Google Scholar]
  2. Chuah M. I., Farbman A. I., Menco B. P. Influence of olfactory bulb on dendritic knob density of rat olfactory receptor neurons in vitro. Brain Res. 1985 Jul 15;338(2):259–266. doi: 10.1016/0006-8993(85)90155-6. [DOI] [PubMed] [Google Scholar]
  3. Chuah M. I., Farbman A. I. Olfactory bulb increases marker protein in olfactory receptor cells. J Neurosci. 1983 Nov;3(11):2197–2205. doi: 10.1523/JNEUROSCI.03-11-02197.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Costanzo R. M., Graziadei P. P. A quantitative analysis of changes in the olfactory epithelium following bulbectomy in hamster. J Comp Neurol. 1983 Apr 20;215(4):370–381. doi: 10.1002/cne.902150403. [DOI] [PubMed] [Google Scholar]
  5. Currier T. C., Nester E. W. Isolation of covalently closed circular DNA of high molecular weight from bacteria. Anal Biochem. 1976 Dec;76(2):431–441. doi: 10.1016/0003-2697(76)90338-9. [DOI] [PubMed] [Google Scholar]
  6. Farbman A. I., Margolis F. L. Olfactory marker protein during ontogeny: immunohistochemical localization. Dev Biol. 1980 Jan;74(1):205–215. doi: 10.1016/0012-1606(80)90062-7. [DOI] [PubMed] [Google Scholar]
  7. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  8. Fitzgerald M., Shenk T. The sequence 5'-AAUAAA-3'forms parts of the recognition site for polyadenylation of late SV40 mRNAs. Cell. 1981 Apr;24(1):251–260. doi: 10.1016/0092-8674(81)90521-3. [DOI] [PubMed] [Google Scholar]
  9. Graziadei G. A., Stanley R. S., Graziadei P. P. The olfactory marker protein in the olfactory system of the mouse during development. Neuroscience. 1980;5(7):1239–1252. doi: 10.1016/0306-4522(80)90197-9. [DOI] [PubMed] [Google Scholar]
  10. Gubler U., Hoffman B. J. A simple and very efficient method for generating cDNA libraries. Gene. 1983 Nov;25(2-3):263–269. doi: 10.1016/0378-1119(83)90230-5. [DOI] [PubMed] [Google Scholar]
  11. Hanahan D., Meselson M. Plasmid screening at high colony density. Gene. 1980 Jun;10(1):63–67. doi: 10.1016/0378-1119(80)90144-4. [DOI] [PubMed] [Google Scholar]
  12. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  13. Kozak M. Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res. 1984 Jan 25;12(2):857–872. doi: 10.1093/nar/12.2.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Margolis F. L. Solid-phase radioimmune assay. Using 3 H-labeled antigen for the mouse olfactory bulb specific protein. Anal Biochem. 1972 Dec;50(2):602–607. doi: 10.1016/0003-2697(72)90071-1. [DOI] [PubMed] [Google Scholar]
  15. Margolis F. L., Tarnoff J. F. Site of biosynthesis of the mouse brain olfactory bulb protein. J Biol Chem. 1973 Jan 25;248(2):451–455. [PubMed] [Google Scholar]
  16. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  17. Menco B. P., Farbman A. I. Genesis of cilia and microvilli of rat nasal epithelia during pre-natal development. I. Olfactory epithelium, qualitative studies. J Cell Sci. 1985 Oct;78:283–310. doi: 10.1242/jcs.78.1.283. [DOI] [PubMed] [Google Scholar]
  18. Milner R. J., Sutcliffe J. G. Gene expression in rat brain. Nucleic Acids Res. 1983 Aug 25;11(16):5497–5520. doi: 10.1093/nar/11.16.5497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Miragall F., Monti Graziadei G. A. Experimental studies on the olfactory marker protein. II. Appearance of the olfactory marker protein during differentiation of the olfactory sensory neurons of mouse: an immunohistochemical and autoradiographic study. Brain Res. 1982 May 6;239(1):245–250. doi: 10.1016/0006-8993(82)90846-0. [DOI] [PubMed] [Google Scholar]
  20. Parnes J. R., Velan B., Felsenfeld A., Ramanathan L., Ferrini U., Appella E., Seidman J. G. Mouse beta 2-microglobulin cDNA clones: a screening procedure for cDNA clones corresponding to rare mRNAs. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2253–2257. doi: 10.1073/pnas.78.4.2253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Rogers K. E., Grillo M., Sydor W., Poonian M., Margolis F. L. Olfactory neuron-specific protein is translated from a large poly(A)+ mRNA. Proc Natl Acad Sci U S A. 1985 Aug;82(15):5218–5222. doi: 10.1073/pnas.82.15.5218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sydor W., Teitelbaum Z., Blacher R., Sun S., Benz W., Margolis F. L. Amino acid sequence of a unique neuronal protein: rat olfactory marker protein. Arch Biochem Biophys. 1986 Sep;249(2):351–362. doi: 10.1016/0003-9861(86)90011-1. [DOI] [PubMed] [Google Scholar]
  23. White B. A., Bancroft F. C. Cytoplasmic dot hybridization. Simple analysis of relative mRNA levels in multiple small cell or tissue samples. J Biol Chem. 1982 Aug 10;257(15):8569–8572. [PubMed] [Google Scholar]

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