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Molecular characterization of a new synthetic cry2ab gene in Nicotiana tabacum
Authors:Sohail Mehmood Karimi  Muhammad Nouman Sohail  Imran Amin  Shahid Mansoor  Zahid Mukhtar
Affiliation:1. National Institute for Biotechnology and Genetic Engineering, P.O. Box 577, Jhang Road, Faisalabad, Pakistan
2. Institute of Molecular Biology and Biotechnology (IMBB), The University of Lahore, Lahore, Pakistan
Abstract:A newly-synthesized cry2Ab gene was characterized in Nicotiana tabacum, before its further transformation in cotton. Synthetic cry2Ab gene was cloned in pGreen0029 and its expression was transiently analyzed at mRNA level through agroinfiltration in tobacco. The mRNA of cry2Ab was detected after 72 h agroinfiltration through PCR using total plant RNA. This construct was then transformed into N. tabacum through Agrobacterium. Insect bioassays were conducted on detached leaves using first instar Spodoptera exigua larvae; after 96 h significant insect mortality was recorded. This newly synthesized gene was effective in controlling S. exigua first instar larvae. It can be used in combinations with other Bt genes like cry1Ac for developing resistance against major insect pests of cotton and further widening the insect control spectrum.
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