Molecular cloning of thioredoxin reductase isozymes, <i>TrxR1</i> and <i>TrxR2</i>, from human lung adenocarcinoma cell line, NCI-H441

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  • Tamura Takashi
    Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University
  • Hasegawa Masaya
    Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University
  • Sugimoto Manabu
    Research Institutes of Bioresources, Okayama University
  • lnagaki Kenji
    Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University
  • Tanaka Hidehiko
    Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University

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  • Molecular cloning of thioredoxin reductase isozymes, TrxR1 and TrxR2, from human lung adenocarcinoma cell line, NCI-H441

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Abstract

<p>The genes encoding mammalian thioredoxin reductase (TrxR) isozymes, TrxR1 and TrxR2, were cloned from a human lung adenocarcinoma cell line, NCI-H441. TrxR1 gene was amplified by conventional thermal cycle program to give an intensive signal on agarose gel electrophoresis. mRNA from normal lung cells also produced the amplified TrxR1 gene by the same RT-PCR procedure. In contrast, the amplification of TrxR2 gene from the NCI-H441 cells required the touch-down PCR program in which the annealing temperature was decreased from 75 to 65 degree by 0.4 degree in every cycle. The normal lung mRNA sample failed to yield the TrxR2 gene, suggesting even smaller expression in the normal cells. The amplified genes were cloned on TOPO TA vector and sequenced to identify the sequences.</p>

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