Skip to main content
SpringerLink
Log in

Construction of a human heart cDNA library and identification of cardiovascular based genes (CVBest)

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

The availability of high quality cDNA libraries is often crucial to the successful identification and characterization of genes. The concepts and potential pitfalls of constructing cDNA libraries are presented. Various applications requiring high quality cDNA libraries are outlined, including large-scale single pass sequencing of cDNA clones to generate expressed sequence tags (ESTs) and differential screening of cDNA libraries. The usefulness of combining such approaches for the discovery of novel disease-related and cardiovascular-based ESTs (CVBest) is discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Chomczynski P, Sacchi N: Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159, 1987

    Google Scholar 

  2. Gubler U, Hoffman BJ: A simple and very efficient method for generating cDNA libraries. Gene 25: 263–269, 1983

    Google Scholar 

  3. D'Alessio JM, Noon MC, Ley III HL, Gerard GF: One-tube doublestranded cDNA synthesis using cloned m-mlv reverse transcriptase. Focus 9: 1–4, 1987

    Google Scholar 

  4. Liew CC: A human heart cDNA library–the development of an efficient and simple method for automated DNA sequencing. J Mol Cell Cardiol 25: 891–894, 1993

    Google Scholar 

  5. Hwang DM, Hwang WS, Liew CC: Single pass sequencing of a unidirectional human fetal heart cDNA library to discover novel genes of the cardiovascular system. J Mol Cell Cardiol 26: 1329–1333, 1994

    Google Scholar 

  6. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ: Basic local alignment search tool. J Mol Biol 215: 403–410, 1990

    Google Scholar 

  7. Gish W, States DJ: Identification of protein coding regions by database similarity search. Nature Genet 3: 266–272, 1993

    Google Scholar 

  8. Liew CC, Hwang DM, Fung YW, Laurenssen C, Cukerman E, Tsui S, Lee CY: A catalogue of genes in the cardiovascular system as identified by expressed sequence tags (ESTs). Proc Natl Acad Sci USA 91: 10645–10649, 1994

    Google Scholar 

  9. Hwang DM, Fung YW, Wang RX, Laurenssen CM, Ng SH, Lam WY, Tsui KW, Fung KP, Waye M, Lee CY, Liew CC: Analysis of expressed sequence tags from a fetal human heart cDNA library. Genomics 30: 293–298, 1995

    Google Scholar 

  10. Fung YW, Liew CC: Identification of genes associated with myocardial development. J Mol Cell Cardiol, in press

  11. Adams MD, Kelley JM, Gocayne JD, Dubnick M, Polymeropoulos MH, Xiao H, Merril CR, Wu A, Olde B, Moreno RF, Kerlavage AR, McCombie WR, Venter JC: Complementary DNA sequencing: expressed sequence tags and human genome project. Science 252: 1651–1656, 1991

    Google Scholar 

  12. Adams MD, Dubnick M, Kerlavage AR, Moreno R, Kelley KT, Utterback TR, Nagle JW, Fields C, Venter JC: Sequence identification of 2,375 human brain genes. Nature 355: 632–634, 1992

    Google Scholar 

  13. Adams MD, Kerlavage AR, Fields C, Venter JC: 3,400 new expressed sequence tags identify a diversity of transcripts in human brain. Nature Genet 4: 256–267, 1993

    Google Scholar 

  14. Adams MD, Soares MB, Kerlavage AR, Fields C, Venter JC: Rapid cDNA sequencing (expressed sequence tags) from a directionally cloned human infant brain cDNA library. Nature Genet 4: 373–380, 1993

    Google Scholar 

  15. Frigerio JM, Berthezene P, Garrido P, Ortiz E, Barthellemy S, Vasseur S, Sastre B, Slelznieff I, Dagorn JC, Iovanna JL: Analysis of 2166 clones from a human colorectal cDNA library by partial sequencing. Hum Mol Genet 4: 37–43, 1995

    Google Scholar 

  16. Gieser L, Swaroop A: Expressed sequence tags and chromosomal localization of cDNA clones from a subtracted retinal pigment epithelium library. Genomics 13: 873–876, 1992

    Google Scholar 

  17. Khan AS, Wilcox AS, Polymeropoulos MH, Hopkins JA, Stevens TJ, Robinson M, Orpana AK, Sikela JM: Single pass sequencing and physical and genetic mapping of human brain cDNAs. Nature Genet 2: 180–185, 1992

    Google Scholar 

  18. Okubo K, Hori N, Matoba R, Niiyama T, Fukushima A, Kojima Y, Matsubara K: Large scale cDNA sequencing for analysis of quantitative and qualitative aspects of gene expression. Nature Genet 2: 173–179, 1992

    Google Scholar 

  19. Sudo K, Chinen K, Nakamura Y: 2058 Expressed sequence tags (ESTs) from a human fetal lung cDNA library. Genomics 24: 276–279, 1994

    Google Scholar 

  20. Takeda J, Yano H, Eng S, Zeng Y, Bell GI: A molecular inventory of human pancreatic islets: sequence analysis of 1000 cDNA clones. Hum Mol Genet 2: 1793–1798, 1993

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. The Cardiac Gene Unit, Departments of Clinical Biochemistry and Medicine, The Centre for Cardiovascular Research, The Toronto Hospital, University of Toronto, Toronto, Ontario, Canada

    C.C. Liew, D.M. Hwang, R.X. Wang, S.H. Ng, A. Dempsey, D.H.Y. Wen, H. Ma, E. Cukerman, X.G. Zhao, Y.Q. Liu, X.K. Qiu, X.M. Zhou & J.R. Gu

  2. The Shanghai Cancer Institute, Shanghai, China

    X.K. Qiu & X.M. Zhou

  3. Department of Biochemistry, The Chinese University of Hong Kong, Shatin, Hong Kong

    C.C. Liew, S. Tsui, K.P. Fung, M.M.W. Waye & C.Y. Lee

Authors
  1. C.C. Liew
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. D.M. Hwang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. R.X. Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. S.H. Ng
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. A. Dempsey
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. D.H.Y. Wen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. H. Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. E. Cukerman
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. X.G. Zhao
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Y.Q. Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. X.K. Qiu
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. X.M. Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. J.R. Gu
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. S. Tsui
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. K.P. Fung
    View author publications

    You can also search for this author in PubMed Google Scholar

  16. M.M.W. Waye
    View author publications

    You can also search for this author in PubMed Google Scholar

  17. C.Y. Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Liew, C., Hwang, D., Wang, R. et al. Construction of a human heart cDNA library and identification of cardiovascular based genes (CVBest). Mol Cell Biochem 172, 81–87 (1997). https://doi.org/10.1023/A:1006811403996

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006811403996

Search

Navigation