Scientific Research and Essays

  • Abbreviation: Sci. Res. Essays
  • Language: English
  • ISSN: 1992-2248
  • DOI: 10.5897/SRE
  • Start Year: 2006
  • Published Articles: 2754

Full Length Research Paper

Cloning and expression of tissue plasminogen activator (t-pa) gene in tobacco plants

  A. Masoumiasl1*, M. Jalali-Javaran1, F. Mahboudi2 and H. Alizadeh3        
  1Department of Plant Breeding, School of Agriculture, Tarbiat Modares University, Tehran, Iran. 2Pasteur Institute of Iran, Tehran, Iran. 3Department of Plant Breeding, School of Agriculture, Tehran University, Tehran, Iran.
Email: [email protected]

  •  Accepted: 18 March 2010
  •  Published: 31 May 2010

Abstract

 

Plants offer a promising alternative to microbial fermentation and animal cell cultures for production of recombinant proteins, the major advantages are safety, low cost, post-translation modifications and high volume of production. t-PA (tissue Plasminogen activator) is a trypsin-like serine proteinase and a superior thrombolytic agent. In this report, recombinant cDNA of tissue Plasminogen activator was transformed to tobacco plants. t-PA recombinant protein was expressed under the control of CaMV35S promoter and NOS terminator. A high-expression sequence (Kozak sequence) and KDEL signal (for endoplasmic reticulum retention of recombinant protein) were linked to amino and carboxy-termini of t-PA gene, respectively. The vector containing t-PA (pBIt-PA) was transferred to Agrobacterium tumefaciens and the t-PA gene was inserted into the plant genome by agrobacterium-mediated transformation. Transgenic plants were selected on kanamycin (100 mgL-1), maintained in perlite and then the soil, subsequent generations were obtained. The presence and expression of the transgene was confirmed in the transformants by PCR, SDS-PAGE, RT-PCR, Zymography and Western blotting. This report examines the transformation and expression of t-PA gene in tobacco plants.

 

Key words: Recombinant protein, tissue plasminogen activator, tobacco, zymography.