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University of Calcutta (U of C), Kolkata

PI: Dr Karabi Datta 

Low phytate rice development: 

To develop low phytate rice, appA gene was cloned from Escherichia coli genomic DNA. Embryo and endosperm specific Zein promoter was cloned from maize (Zea mays) genomic DNA. Over-expression vector was constructed with appA gene under the control of zein promoter. Seven low phytate transgenic lines were developed showing 30-50% reduction in phytic acid level. Mineral content of transgenic lines were significantly higher than that of the non-transgenic control.

 

 

Low phytate rice:

Over expression of E.coli phytase gene 

 

 

 

 

 

 

 

 

 

 

 Lysine content increment:

Two approaches are being attempted viz., Seed specific expression of less feedback sensitive bacterial DHDPS (dihydrodipicolinate synthase) enzyme  and RNAi mediated downregulation of bifunctional lysine degrading LKR/SDH (lysine ketoglutarate reductase/saccharopine dehydrogenase) enzyme.  The first approach for lysine content increment in rice, DHDPS gene from Corynebacterium glutamicum was cloned under zein promoter and rice transformation was done with the vector. Four lines were initially selected by molecular analysis. Analysis of amino acid content reveals that lysine content has been improved in transgenic seeds by1.5 times of that of the WT).For the second approach, lkr/sdh gene fragment (452bp) was isolated from cDNA of rice (Oryza sativa) and RNAi vector was constructed with glutelinC promoter from rice in destination vector. Rice transformation was performed by Agrobacterium mediated transformation method. Analysis of amino acid content reveals that lysine content is improved in transgenic seeds of the down regulation lines (2.6 times of that of the WT). 



 

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