title: Expression analysis and purification of human recombinant tissue type plasminogen activator (rt-PA) from transgenic tobacco plants.
abstract: Recombinant tissue-type plasminogen activator (rt-PA) has been produced in different hosts. In this research, transgenic tobacco was selected for production of human rt-PA. Transgenic plants were analyzed by polymerase chain reaction (PCR) and reverse-transcription (RT)-PCR. The protein was extracted by Lysine Sepharose chromatography column and was further purified by HiTrap desalting column. The function of eluted protein was analyzed on zymography gel. The results showed that the 1.7-kb cDNA of tissue-type plasminogen activator (t-PA) (as well as a shortened 650-bp transcript of t-PA) has been expressed in transgenic plants. The anticipated 63-kD protein band and an additional 53-kD protein were observed in transgenic plants. Finally, zymography assay revealed that the purified rt-PA has anticipated appropriate activity comparable to a positive control drug (Alteplase). On the whole, we can say that transgenic tobacco is a good alternative host for production of t-PA.
authors: Department of Biotechnology, School of Agriculture, Tarbiat Modares University, Tehran, Iran.
authors: Haidar Saify
authors: HS
authors: Nabiabad
authors: Mohammad Mehdi
authors: MM
authors: Yaghoobi
authors: Mokhtar Jalali
authors: MJ
authors: Javaran
authors: Saman
authors: S
authors: Hosseinkhani
mesh_terms: D016960:Agrobacterium tumefaciens; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D005341:Fibrinolysin / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D010958:Plasminogen / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D010959:Tissue Plasminogen Activator / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification*; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D010958:Plasminogen; D010959:Tissue Plasminogen Activator; D005341:Fibrinolysin