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Cloning and chloroplast-targeted expression studies of insect-resistant gene with ricin fusion-gene under chloroplast transit peptide in cotton
Kiani, Sarfraz; Ali, Arfan; Bajwa, Kamran Shehzad; Muzaffar, Adnan; Ashraf, Muhammad Aleem; Samiullah, Tahir Rehman; Shahid, Ahmad Ali; Husnain, Tayyab.
Afiliación
  • Kiani, Sarfraz; University of the Punjab. National Centre of Excellence in Molecular Biology. Lahore. PK
  • Ali, Arfan; University of the Punjab. National Centre of Excellence in Molecular Biology. Lahore. PK
  • Bajwa, Kamran Shehzad; University of the Punjab. National Centre of Excellence in Molecular Biology. Lahore. PK
  • Muzaffar, Adnan; University of the Punjab. National Centre of Excellence in Molecular Biology. Lahore. PK
  • Ashraf, Muhammad Aleem; University of the Punjab. National Centre of Excellence in Molecular Biology. Lahore. PK
  • Samiullah, Tahir Rehman; University of the Punjab. National Centre of Excellence in Molecular Biology. Lahore. PK
  • Shahid, Ahmad Ali; University of the Punjab. National Centre of Excellence in Molecular Biology. Lahore. PK
  • Husnain, Tayyab; University of the Punjab. National Centre of Excellence in Molecular Biology. Lahore. PK
Electron. j. biotechnol ; Electron. j. biotechnol;16(6): 13-13, Nov. 2013. ilus, tab
Article en En | LILACS | ID: lil-696554
Biblioteca responsable: CL1.1
ABSTRACT

Background:

Transgenic plants inhabiting single Bt gene are prone to develop insect resistance and this resistance has been reported in case of some important yield-devastating insect larvae of commercial crops, such as cotton and rice. Therefore, it has become essential to adapt new strategies to overcome the problem of insect resistance and these new strategies should be sophisticated enough to target such resistant larvae in broad spectrum. Among these, plants may be transformed with Bt gene tagged with some fusion-protein gene that possesses lectin-binding capability to boost the binding sites for crystal protein gene within insect mid-gut in order to overcome any chances of insect tolerance against Bt toxin. Enhanced chloroplast-targeted Bt gene expression can also help in the reduction of insect resistance.

Results:

In the present investigation, a combined effect of both these strategies was successfully used in cotton (G. hirsutum). For this purpose, plant expression vector pKian-1 was created, after a series of cloning steps, carrying Cry1Ac gene ligated with chloroplast transit peptide towards N-terminal and Ricin B-Chain towards C-terminal, generating TP-Cry1Ac-RB construct.

Conclusions:

Efficacy of pKian-1 plasmid vector was confirmed by in-planta Agrobacterium-mediated leaf GUS assay in tobacco. Cotton (G. hirsutum) local variety MNH-786 was transformed with pKian-1 and the stable integration of TP-Cry1Ac-RB construct in putative transgenic plants was confirmed by PCR; while fusion-protein expression in cytosol as well as chloroplast was substantiated by Western blot analysis. Whereas, confocal microscopy of leaf-sections of transgenic plants exposed that hybrid-Bt protein was expressing inside chloroplasts.
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Texto completo: 1 Colección: 01-internacional Base de datos: LILACS Asunto principal: Cloroplastos / Plantas Modificadas Genéticamente / Proteínas de Cloroplastos Idioma: En Revista: Electron. j. biotechnol Asunto de la revista: BIOTECNOLOGIA Año: 2013 Tipo del documento: Article País de afiliación: Pakistán Pais de publicación: Chile
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: LILACS Asunto principal: Cloroplastos / Plantas Modificadas Genéticamente / Proteínas de Cloroplastos Idioma: En Revista: Electron. j. biotechnol Asunto de la revista: BIOTECNOLOGIA Año: 2013 Tipo del documento: Article País de afiliación: Pakistán Pais de publicación: Chile