Cloning of PA1 gene from Pisum sativum and genetic transformation on Medicago sativa

Abstract

Abstract: A gene, a sulphur-rich and insect-toxic seed albumin-PA1 gene, was cloned by PCR method from Pisum sativum (cultivar shijiadacaiwan). In order to characterize the function of this gene, a plant expression vector, pCAMBIA1301-PA1, was constructed. This vector was transformed into M. sativa via Agrobacterium-mediated method. The conditions for transformation were optimized. The several transgenic embryogenic calli and regenerated plantlets were obtained. PCR and Southern blotting identification confirmed that PA1 gene and hygromycin-B phosphotransferase-resistant gene has been integrated into the genome of M. sativa, SDS-PAGE analysis indicated that PA1 gene expressed in the transgenic lines. Free amino acid analysis showed that the levels of methionine and cysteine in tansgenic M. sativa. Plantlets improved from 0.1% to 0.4%.

CLC Number:

ZHANG Gai-na, JIA Jing-fen. Cloning of PA1 gene from Pisum sativum and genetic transformation on Medicago sativa[J]. Acta Prataculturae Sinica, 2009, 18(3): 117-125.

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