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Cloning Genes From an Inner Ear cDNA Library

Cloning Genes From an Inner Ear cDNA Library Abstract A rat inner ear complementary DNA (cDNA) library containing 1.9 ×106 recombinants was constructed and evaluated. Inserts averaged 2.0 (±2.1) kilobases in length. A subset of inserts was screened for site of expression. Two cDNA transcripts were isolated based on cochlear expression restricted to the spiral ganglion. One transcript showed a high degree of homology to several long interspersed DNA elements, neuron-specific nuclear transscripts thought to be involved in gene regulation. The second transcript showed no homology to known sequences and appears to encode a neuron-specific protein of about 248 amino acids. The library can be used to identify proteins important for inner ear function and disease. (Arch Otolaryngol Head Neck Surg. 1993;119:1217-1220) References 1. Huynh T, Young R, Davis R. Construction and screening cDNA libraries in lambda gt10 and lambda gt11 . In: Glover DM, ed. DNA Cloning: A Practical Approach . Washington, DC: IRL Press; 1985;1:49-78. 2. Simmons DM, Arriza JL, Swanson LW. A complete protocol for in situ hybridization of messenger RNAs in brain and other tissues with radiolabeled single-stranded RNA probes . J Histotechnol . 1989;12:169-181.Crossref 3. Ryan AF, Watts AG, Simmons DM. The preservation of mRNA during in situ hybridization in the cochlea . Hearing Res . 1991;56:148-152.Crossref 4. Sanger F, Nicklen S, Coulson AR. DNA sequence with chain-terminating inhibitors . Proc Natl Acad Sci U S A . 1977;74:5463-5467.Crossref 5. Fickett JW. Recognition of protein coding regions in DNA sequences . Nucl Acids Res . 1982;10:5303-5318.Crossref 6. Ryan AF. The relationship between metabolism and blood flow in the cochlea . In: Kahn S, Santos-Sacchi J, eds. Physiology of the Ear . New York, NY: Raven Press; 1988:317-326. 7. Ryan AF, Watts AG. Expression of genes coding for α and β isoforms of Na/K-ATPase in the cochlea of the rat . Cell Mol Neurosci . 1991;2:179-187.Crossref 8. D'Ambrosio E, Waitzkin SD, Witney FR, Salemme A, Furano AV. Structure of the highly repeated, long interspersed DNA family (LINE or L1Rn) of the rat . Mol Cell Biol . 1986;6:411-424. 9. Schmitz E, Mohr E, Richter D. Rat vasopressin and oxytocin genes are linked by a long interspersed repeated DNA element (LINE): sequence and transcriptional analysis on LINE . DNA Cell Biol . 1991;10:81-91.Crossref 10. Lewin B. Genes IV . Cambridge, Mass: Cell Press; 1990. 11. Wilcox E, Fex J. Construction of a cDNA library from microdissected guinea pig organ of Corti . Hearing Res . 1992;62:124-126.Crossref 12. Ausubel FM, Brent R, Kingston RE, et al, eds. Current Protocols in Molecular Biology . New York, NY: John Wiley & Sons; 1990;1:5.5.8-13. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Otolaryngology - Head & Neck Surgery American Medical Association

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Publisher
American Medical Association
Copyright
Copyright © 1993 American Medical Association. All Rights Reserved.
ISSN
0886-4470
eISSN
1538-361X
DOI
10.1001/archotol.1993.01880230059010
Publisher site
See Article on Publisher Site

Abstract

Abstract A rat inner ear complementary DNA (cDNA) library containing 1.9 ×106 recombinants was constructed and evaluated. Inserts averaged 2.0 (±2.1) kilobases in length. A subset of inserts was screened for site of expression. Two cDNA transcripts were isolated based on cochlear expression restricted to the spiral ganglion. One transcript showed a high degree of homology to several long interspersed DNA elements, neuron-specific nuclear transscripts thought to be involved in gene regulation. The second transcript showed no homology to known sequences and appears to encode a neuron-specific protein of about 248 amino acids. The library can be used to identify proteins important for inner ear function and disease. (Arch Otolaryngol Head Neck Surg. 1993;119:1217-1220) References 1. Huynh T, Young R, Davis R. Construction and screening cDNA libraries in lambda gt10 and lambda gt11 . In: Glover DM, ed. DNA Cloning: A Practical Approach . Washington, DC: IRL Press; 1985;1:49-78. 2. Simmons DM, Arriza JL, Swanson LW. A complete protocol for in situ hybridization of messenger RNAs in brain and other tissues with radiolabeled single-stranded RNA probes . J Histotechnol . 1989;12:169-181.Crossref 3. Ryan AF, Watts AG, Simmons DM. The preservation of mRNA during in situ hybridization in the cochlea . Hearing Res . 1991;56:148-152.Crossref 4. Sanger F, Nicklen S, Coulson AR. DNA sequence with chain-terminating inhibitors . Proc Natl Acad Sci U S A . 1977;74:5463-5467.Crossref 5. Fickett JW. Recognition of protein coding regions in DNA sequences . Nucl Acids Res . 1982;10:5303-5318.Crossref 6. Ryan AF. The relationship between metabolism and blood flow in the cochlea . In: Kahn S, Santos-Sacchi J, eds. Physiology of the Ear . New York, NY: Raven Press; 1988:317-326. 7. Ryan AF, Watts AG. Expression of genes coding for α and β isoforms of Na/K-ATPase in the cochlea of the rat . Cell Mol Neurosci . 1991;2:179-187.Crossref 8. D'Ambrosio E, Waitzkin SD, Witney FR, Salemme A, Furano AV. Structure of the highly repeated, long interspersed DNA family (LINE or L1Rn) of the rat . Mol Cell Biol . 1986;6:411-424. 9. Schmitz E, Mohr E, Richter D. Rat vasopressin and oxytocin genes are linked by a long interspersed repeated DNA element (LINE): sequence and transcriptional analysis on LINE . DNA Cell Biol . 1991;10:81-91.Crossref 10. Lewin B. Genes IV . Cambridge, Mass: Cell Press; 1990. 11. Wilcox E, Fex J. Construction of a cDNA library from microdissected guinea pig organ of Corti . Hearing Res . 1992;62:124-126.Crossref 12. Ausubel FM, Brent R, Kingston RE, et al, eds. Current Protocols in Molecular Biology . New York, NY: John Wiley & Sons; 1990;1:5.5.8-13.

Journal

Archives of Otolaryngology - Head & Neck SurgeryAmerican Medical Association

Published: Nov 1, 1993

References