Article

Biotechnology Letters

, Volume 15, Issue 11, pp 1095-1100

Molecular cloning and functional expression of a Talaromyces emersonii derived alpha-amylase encoding genetic determinant in a human cell line

  • L. BunniAffiliated withDept of Biological and Biomedical Sciences, University of UlsterDept of Microbiology, Trinity College
  • , T. J. HackettAffiliated withDept of Biological and Biomedical Sciences, University of UlsterArthur Anderson Consulting
  • , L. McHaleAffiliated withDept of Biological and Biomedical Sciences, University of Ulster
  • , G. FlynnAffiliated withDept of Biological and Biomedical Sciences, University of Ulster
  • , A. P. McHaleAffiliated withDept of Biological and Biomedical Sciences, University of Ulster

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Summary

A cDNA fragment encoding a Talaromyces emersonii acid stable alpha-amylase has been cloned into a mammalian cell expression vector system. When human HeLa cells were transformed with this DNA, functional enzyme could be detected in extracellular media and cell lysates derived from stable transformants. Expression of the T. emersonii derived cDNA was verified by northern dot blotting hybridisation analysis of mRNA extracted from transformants, using the labelled amylase encoding cDNA determinant as a probe. Results obtained suggest that expression of the fungus derived alpha-amylase in the transformed HeLa cells is glucocorticoid dependent.