Transformasi Genetik Tembakau dengan Gen Cold Shock Protein melalui Perantara Agrobacterium tumefaciens

Seagames Waluyo, Sustiprijatno Sustiprijatno, Suharsono Suharsono

Abstract


Cold
shock protein (Csp) essential for organisms to survive in
abiotic stress condition. CspB gene has been fused to
ubiquitin promoter in the T-DNA region of pCambia 1300int,
and introduced into Agrobacterium tumefaciens LBA4404.
This research had an objective to transform genetically
Nicotiana tabacum cv. Samsun by CspB gene under the
control of Ubiquitin promoter and NOS terminator mediated
by A. tumefaciens. Leaf discs were co-cultivated with A.
tumefaciens LBA 4404. Based on the number of hygromycinresistant
calli, the efficiency of transformation was 57.5%. In
the selective medium containing 50 μg/l hygromycin, the
efficiency of regeneration of transgenic shoots was 82.6%.
Based on PCR analysis using primers corresponding to
ubiquitin promoter and CspB gene, 18 putative tobacco
transgenic containing CspB gene under the control of
ubiquitin promoter.


Keywords


Tobacco (Nicotiana tabaccum); Cold Shock Protein (Csp); transformation; Agrobacterium tumefaciens.

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DOI: http://dx.doi.org/10.21082/jbio.v9n2.2013.p58-65

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P-ISSN : 1907-1094
E-ISSN : 2549-1547


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