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Analysis of the in Vitro Translation Product of mRNA Coding for Chick Intestine Apolipoprotein A-I

  • S. Ferrari
  • E. Drusiani
  • P. Tarugi
  • S. Calandra
Part of the GWUMC Department of Biochemistry Annual Spring Symposia book series (GWUN)

Abstract

In all animal species studied so far. apolipoprotein A-I, the main constituent peptide of plasma HDL, is present in plasma as a family of isoforms that have the same molecular weight but different isoelectric points. After in vitro translation of chick intestine mRNA, analysis of the primary translation product of immunoprecipitated apo A-I, by two-dimensional gel electrophoresis, also reveals the presence of at least three isoforms. The origin of these isoforms is still unclear. It may reflect the presence of different mRNAs, each encoding a single apo A-I isoform, or derive from posttranslational modifications of a single translation product. To answer this question we first isolated a “full-size” cDNA representing the coding sequence of chick intestinal mRNA by screening a cDNA library in pEX-1 expression vector. The cDNA insert was then subcloned in p-GEM 1 vector and transcribed with SP 6 polymerase. This in vitro transcribed RNA was then translated in a cell-free system in the presence of [35S]methionine. The primary translation product was immunoprecipitated with anti-apo A-I antiserum and subjected to two-dimensional gel electrophoresis. The isoform pattern of this translation product was found to be identical to that observed after translation of total RNA extracted from chick intestinal mucosa. This finding indicates that the presence of multiple isoforms in the primary translation product of chick intestine apo A-I appears to result from posttranslational modification of a single translation product.

Keywords

Translation Product cDNA Insert Multiple Isoforms mRNA Code Primary Translation Product 
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

© Plenum Press, New York 1987

Authors and Affiliations

  • S. Ferrari
    • 1
  • E. Drusiani
    • 1
  • P. Tarugi
    • 2
  • S. Calandra
    • 2
  1. 1.Istituto di Chimica BiologicaUniversità di ModenaModenaItaly
  2. 2.Istituto di Patologia GeneraleUniversità di ModenaModenaItaly

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