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TRANSFER AND EXPRESSION OF RABBIT DEFENSIN GENE NP-1 IN CUMCUMBER. Song Dan, Zhou Wenting, Qian Wenjing, Liu Jing, He Zhengquan

TRANSFER AND EXPRESSION OF RABBIT DEFENSIN GENE NP-1 IN CUMCUMBER.

Song Dan, Zhou Wenting, Qian Wenjing, Liu Jing, He Zhengquan

International Journal of Biochemistry & Biotech Sciences 2013;2(1): 1-10

Abstract

The rabbit defensin NP-1 is one of the defensins so far showed the broadest spectrum of antimicrobial and strong antibacterial activity, which mainly expressed in rabbit neutrophil. The rabbit defensin NP-1 has been transferred into cucumber to study the treatment role on human gastric disease. Plasmid pBinU-Ome-HisNP1 was introduced into cucumber using Agrobacterium strain LBA4404. Transgenic cucumber plants expressing rabbit defensin gene (NP-1) have been successfully selected and regenerated on MS medium supplemented with kalamycin (25 mg l-1), 0.05 mg l-1 6-benzylaminopurine and 0.1 mg l-1 indolebutyric acid. Integration of the NP-1 gene into cucumber genome was confirmed by polymerase chain reaction (PCR), Southern blot, Westhern blot analysis and resistance tests. PCR amplified a 130bp characteristics fragment corresponds to the fragment from coding region of the defensin NP-1 mature peptide. Southern blot analysis confirmed the integration of 2.3 kb NP-1 gene, while Western blot analysis detected the presence of 42KD protein. It was demonstrated that the target gene expressed in cucumber preliminarily. Resistance tests demonstrated that the expression of native defensin NP-1 could confer partial resistance to E. coil and Staphylococcus aureus, which have made immense progress in cucumber disease resistance and breeding.

Keywords: Rabbit defensin (NP-1) gene, cucumber, transformation.
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