Author/Authors :
Durrani, Irfan Safdar University of Agriculture - Institute of Biotechnology and Genetic Engineering, Pakistan , Bakht, Jehan University of Agriculture - Institute of Biotechnology and Genetic Engineering, Pakistan , Swati, Zahoor Ahmad University of Agriculture - Institute of Biotechnology and Genetic Engineering, Pakistan , Naqvi, S.M. Saqlan Pir Mehr Ali Shah Arid Agriculture University (PMAS Arid Agriculture University), Pakistan , Shafi, Mohammad University of Agriculture - Dept of Agronomy, Pakistan
Abstract :
This investigation was carried out to clone rice germin-like protein gene (OsRGLP9) under the transcriptional control of a constitutive promoter in tobacco. For the purpose OsRGLP9 gene, 995 bp was first successfully cloned in pENTRD/Topo® cloning vector and the target gene was sub-cloned in pH7WG2 vector in sense direction downstream of CaMV35S promoter. Presence of insert in the destination vector was confirmed through sequencing. After confirmation recombinant plasmid containing target gene was prepared on large scale by alkaline lysis method and purified using Eppendorf Perfectprep® Gel Cleanup kit. Transgenic plants were obtained through Agro-bacterium mediated transformation. The transgenic plants were confirmed by polymerase chain reaction (PCR).
Keywords :
Cloning , germin like protein , OsRGLP9 , tobacco
