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Journal of Molecular Neuroscience

, Volume 3, Issue 2, pp 59–64 | Cite as

Construction of spinal cord cDNA library and application for subtractive cloning of spinal cord-specific cDNAs

  • H. Kobayashi
  • H. Takahashi
  • K. Oyanagi
  • F. Ikuta
  • T. Miyatake
  • S. Tsuji
Article

Abstract

Neurodegenerative diseases are characterized by neuronal degeneration of specific neurons, e.g., degeneration of motoneurons in amyotrophic lateral sclerosis. As an approach to understand molecular mechanisms of neuronal degeneration of human spinal cord motoneurons in various motor neuron diseases, we have constructed a human spinal cord cDNA library and developed a strategy for isolating spinal cord-specific genes by subtractive cloning. We constructed human spinal cord and brain cDNA libraries from postmortem human spinal cord and brain. To isolate human spinal cord-specific cDNAs, a spinal cord-enriched [32P]cDNA probe was generated by the phenol emulsion reassociation technique. Forty-eight cDNA clones out of 10,000 colonies gave strong signals with the subtracted probe, and individual spinal cord cDNA clones were isolated. Northern blotting analysis confirmed that two spinal cord cDNA clones are, in fact, more abundant in spinal cord compared to brain.

Keywords

Spinal Cord Amyotrophic Lateral Sclerosis cDNA Library Spinal Muscular Atrophy cDNA Probe 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Birkhäuser 1991

Authors and Affiliations

  • H. Kobayashi
    • 1
  • H. Takahashi
    • 2
  • K. Oyanagi
    • 3
  • F. Ikuta
    • 2
  • T. Miyatake
    • 1
  • S. Tsuji
    • 1
  1. 1.Department of NeurologyBrain Research Institute, Niigata UniversityNiigataJapan
  2. 2.Department of PathologyBrain Research Institute, Niigata UniversityNiigataJapan
  3. 3.The Center for Materials of Brain DiseasesBrain Research Institute, Niigata UniversityNiigataJapan

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