Cloning of Tissue Plasminogen Activator cDNA in Nonpathogenic Leishmania

Cell Journal

关键词: Leishmania Tarentolae;    Tissue Plasminogen Activator;    Polymerase Chain Reaction;   

DOI  :  

来源: DOAJ

【 摘 要 】

Introduction: At present, most recombinant proteins are produced inprokaryotes especially E coli. Yeasts and CHO also are used aseukaryotic hosts. Leishmania tarentolae, a parasite of lizards, amember of Trypanosomatidae family is one of the new systems forexpression of heterologous proteins. In this system, some of theparasitic protozoa features are used in expression of mammalianproteins.Material and Methods: For evaluation of the protozoa forexpression of human complex proteins, we cloned cDNA of tPA genecontaining native human signal sequence. We used vectorscontaining 3´ and 5´ sequences of Leishmania 18s rRNA forintegration of the vectors in 18s rRNA gene and severe transcription.Results: RT-PCR test showed production of specific mRNA of tPAgene in the recombinant cells. Southern blot analysis confirmed thecloning of t-PA in the genome of the Leishmania.Conclusion: This study showed native human signal sequencemediate transport and secretion of the protein. Hence, L. tarentolaeis the first useful biotechnologically protozoan and tPA is the mostcomplex protein expressed in it.

【 授权许可】

Unknown