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Construction of pcDNA3.lAM and expression of adrenomedullin in mammalian cells

WANG Xiao- Fang, SHAO Ying, Tian Dezhi, Yao Tai, LU Li- Min

Department of Physiology and Pathaphysiology，Shanghai Medical College，Fudan University，Shanghai 200032

Abstract

The newly discovered endogenous vasodilating and diuretic peptide adrenomedullin(AM)was considered to be of attractive value in clinical treatment of hypertension and congestive heart failure.In order to explore the treatment of cardiovascular diseases by expressing AM in vivo，AM cDNA was
inserted into mammalian expressing vector pcDNA31，and in vitro expression of AM was carried out in cultured K562 cell line AM mRNA was amplified by RT-PCR from the total RNA isolated from the adrenal glands of rats and was inserted into pcDNA31 vector to form pcDNA3I AM，the recombinant
pc DNA31 AM was then transferred into cultured K562 cell line by posome .The expression of AM in pc DNA3.1 AM transferred cell was identified by RT- PCR and dot i m munoblot assay.The results demonstrated that there were AM mRNA in the pcDNA31 ANPtransferred K562 cell line and AM peptides in the culturing medium，indicating that the recombinant pcDNA3 1 AM vector can express AM in mammalian cell line.

Key words: adreno medullin;;K_(562) cell line;

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Citing This Article：

WANG Xiao- Fang, SHAO Ying, Tian Dezhi, Yao Tai, LU Li- Min. Construction of pcDNA3.lAM and expression of adrenomedullin in mammalian cells. Acta Physiol Sin 2003; 55 (1): (in Chinese with English abstract).