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

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

Research output: Contribution to journalArticle

14 Citations (Scopus)

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
StatePublished - Aug 1999

Fingerprint

connexins
Connexins
Screening
reverse transcriptase polymerase chain reaction
screening
Polymerase Chain Reaction
DNA Primers
DNA primers
Oligonucleotides
Tissue
Mammals
gap junctions
Endonucleases
Gap Junctions
protein aggregates
cultured cells
Vertebrates
Cultured Cells
Genes
vertebrates

Keywords

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

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Medicine (miscellaneous)

Cite this

A simple RT-PCR-based strategy for screening connexin identity. / Urban, M.; Rozental, R.; Spray, David C.

In: Brazilian Journal of Medical and Biological Research, Vol. 32, No. 8, 08.1999, p. 1029-1037.

Research output: Contribution to journalArticle

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