Construction of antifreeze gene CBF2 expression vector and transformation into alfalfa callus

Abstract

Abstract: Arabidopsis thaliana genomic DNA was selected as a template to amplify the target gene by PCR and connect it to the PGEM-T Easy Vector for construction of the T-CBF₂. The target fragment and linear plasmids were obtained from the cloning vector T-CBF₂ and from the plant expression vector PBI121 with dual digestion using BamHⅠand Sac I, respectively. The plant expression vector P-T-CBF₂ was built through directional connections using T₄ DNA ligase. PCR identification proved that the recombinant had been transferred into Agrobacterium tumefaciens and it was then introduced into Medicago sativa cv. Hetian through Agrobacterium-mediated transformation. Transfer of the target gene to alfalfa callus was successful.

CLC Number:

LIU Xiao-jing, HAO Feng, ZHANG De-gang, MAO Juan, YU Tie-feng. Construction of antifreeze gene CBF₂ expression vector and transformation into alfalfa callus[J]. Acta Prataculturae Sinica, 2011, 20(2): 193-200.

References

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Construction of antifreeze gene CBF₂ expression vector and transformation into alfalfa callus

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