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Isolation of Unknown Genes from Human Bone Marrow by Differental Screening and Single-Pass cDNA Sequences Determination

Orr, Sally R. and Hughes, Tim P. and Sawyers, Charles L. and Kato, Roberta M. and Quan, Shirley G. and Williams, Suzanne P. and Witte, Owen N. and Hood, Leroy (1994) Isolation of Unknown Genes from Human Bone Marrow by Differental Screening and Single-Pass cDNA Sequences Determination. Proceedings of the National Academy of Sciences of the United States of America, 91 (25). pp. 11869-11873. ISSN 0027-8424. PMCID PMC45337.

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A cDNA sequencing project was initiated to characterize gene expression in human bone marrow and develop strategies to isolate novel genes. Forty-eight random cDNAs from total human bone marrow were subjected to single-pass DNA sequence analysis to determine a limited complexity of mRNAs expressed in the bone marrow. Overall, 8 cDNAs (17%) showed no similarity to known sequences. Information from DNA sequence analysis was used to develop a differential prescreen to subtract unwanted cDNAs and to enrich for unknown cDNAs. Forty-eight cDNAs that were negative with a complex probe were subject to single-pass DNA sequence determination. Of these prescreened cDNAs, the number of unknown sequences increased to 23 (48%). Unknown cDNAs were also characterized by RNA expression analysis using 25 different human leukemic cell lines. Of 13 unknown cDNAs tested, 10 were expressed in all cell types tested and 3 revealed a hematopoietic lineage-restricted expression pattern. Interestingly, while a total of only 96 bone marrow cDNAs were sequenced, 31 of these cDNAs represent sequences from unknown genes and 12 showed significant similarities to sequences in the data bases. One cDNA revealed a significant similarity to a serine/threonine-protein kinase at the amino acid level (56% identity for 123 amino acids) and may represent a previously unknown kinase. Differential screening techniques coupled with single-pass cDNA sequence analysis may prove to be a powerful and simple technique to examine developmental gene expression.

Item Type:Article
Related URLs:
URLURL TypeDescription CentralArticle
Witte, Owen N.0000-0003-4461-4533
Hood, Leroy0000-0001-7158-3678
Additional Information:© 1994 by the National Academy of Sciences. Contributed by Leroy Hood, August 5, 1994. We gratefully acknowledge Mike Fogliano for assistance with the computer analysis. This work was supported by the Lucille P. Markey Charitable Trust (S.L.O.), the Cancer Research Institute, and the Jaye Haddad-Concern Foundation Fellowship (T.P.H.). C.L.S. is a Howard Hughes Medical Institute Research Fellow and O.N.W. is an Investigator of the Howard Hughes Medical Institute. Portions of this work were funded by grants to O.N.W. from the National Cancer Institute. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Funding AgencyGrant Number
Lucille P. Markey Charitable TrustUNSPECIFIED
Cancer Research InstituteUNSPECIFIED
Jaye Haddad-Concern FoundationUNSPECIFIED
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
National Cancer InstituteUNSPECIFIED
Issue or Number:25
PubMed Central ID:PMC45337
Record Number:CaltechAUTHORS:ORRpnas94
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:5198
Deposited By: Archive Administrator
Deposited On:04 Oct 2006
Last Modified:02 Oct 2019 23:20

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