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Transformation and confirmation of GUS gene expression in Solanum melongena L. of PLR 1 cultivar

Vinod Kanna S., Jayabalan N.

Abstract


In the present study GUS gene transformation was carried out in eggplant using Agrobacterium strain with pBAL2 vector harboring gus gene and nptII as selection marker gene. The factors which are affecting (enhancing) the frequency of transient gus gene expression are different physical and biochemical variables has been carried out. It is observed that the 4 day precultured explants showed the minimum survival rate in the medium when compared with 2-day co cultivated medium. The explants which had undergone co-cultivation for 4 to 5 days showed GUS activity, the tissues were adversely affected due to the overgrowth of bacteria. The gene specific primers for nptII and gus gene were used for amplification and it has given 680bp and 1.9 kb amplified fragments respectively and recorded. The band was detected in the selected plants, but it was absent from the negative control (non-transformed) plant in the Southern hybridization. Our experiment showed 0.80-1.60 percentage of efficiency in transformation. With a total of 849 infected shoots were undergone confirmation tests which results 9 PCR positives (1.06% efficiency). The Transformant kept in the Environmental Growth Chamber and transferred to field condition subsequently.

Keywords


Agrobacterium tumifaciens, MS Salts, infection, pBAL2 and transformant

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References


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DOI: https://doi.org/10.21746/aps.2018.7.4.19

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