A simple RT-PCR-based strategy for screening connexin identity

M. Urban, R. Rozental, D. C. Spray

Research output: Contribution to journalArticle

14 Scopus citations

Abstract

Vertebrate gap junctions are aggregates of transmembrane channels which are composed of connexin (Cx) proteins encoded by at least fourteen distinct genes in mammals. Since the same Cx type can be expressed in different tissues and more than one Cx type can be expressed by the same cell, the thorough identification of which connexin is in which cell type and how connexin expression changes after experimental manipulation has become quite laborious. Here we describe an efficient, rapid and simple method by which connexin type(s) can be identified in mammalian tissue and cultured cells using endonuclease cleavage of RT-PCR products generated from "multi primers" (sense primer, degenerate oligonucleotide corresponding to a region of the first extracellular domain; antisense primer, degenerate oligonucleotide complementary to the second extracellular domain) that amplify the cytoplasmic loop regions of all known connexins except Cx36. In addition, we provide sequence information on RT-PCR primers used in our laboratory to screen individual connexins and predictions of extension of the "multi primer" method to several human connexins.

Original languageEnglish (US)
Pages (from-to)1029-1037
Number of pages9
JournalBrazilian Journal of Medical and Biological Research
Volume32
Issue number8
DOIs
StatePublished - Aug 1999

Keywords

  • Cap junctions
  • Intercellular communication
  • Multi primers
  • Universal primers
  • cDNA sequences

ASJC Scopus subject areas

  • Biophysics
  • Neuroscience(all)
  • Biochemistry
  • Physiology
  • Immunology
  • Pharmacology, Toxicology and Pharmaceutics(all)
  • Cell Biology

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