Lymphocyte proteinpaedia stage two: T-cell polypeptides from a partitioned cdna library revealed by the dual decay method

J. R. Kettman, C. Coleclough, I. Lefkovits

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

A complex population of gene products was analyzed by combining the great resolving power of two-dimensional (2D) protein electrophoresis with a de-tailed dissection of individual protein species afforded by cDNA cloning. A cDNA library from BW5147 was partitioned by random sampling into sectors (of a complexity of 500 phase plaques/sector). From each of the unique sets of cDNA clones present in the sector, 2D gels were prepared. Patterns of spots were analyzed using the Kepler software system, and the compiled data both from the sectors and from the natural lymphocyte populations have been established to serve as a guide for gene retrieval. Use of the dual decay method, which compares two exposures of a 2D gel co-electrophoretic pattern, obtained from samples with (35S)-polypeptides mixed with (3H)-polypeptides, is described and scrutinized. It was determined that of 268 spots (three sectors combined), 118spots were shared with the natural lymphocyte population, the remaining 150 spots occur only in the sectors (and not detected in the cell population). All cDNA populations are available for retrieval. Information obtained throughout the work was entered into the database.

Original languageEnglish (US)
Pages (from-to)131-142
Number of pages12
JournalInternational Archives of Allergy and Immunology
Volume103
Issue number2
DOIs
StatePublished - Jan 1 1994

    Fingerprint

Keywords

  • CDNA library
  • Gels
  • Gene catalog
  • Lymphocytes
  • Proteinpaedia
  • Two-dimensional

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

Cite this