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Bulletin of Botanical Research ›› 2008, Vol. 28 ›› Issue (5): 564-567.doi: 10.7525/j.issn.1673-5102.2008.05.013

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The Transformation of Insect Antifreezing Protein for Sauropus androgynus Mediated by Agrobacterium

XIAO Yan-Hua;ZENG You-Ling;LAN Hai-Yan;ZHANG Fu-Chun*   

  1. (Xinjiang Key Laboratory of Biological Resources and Genetic Engineering,College of Life Science and Technology,Xinjiang University,Urumqi 830046)
  • Received:1900-01-01 Revised:1900-01-01 Online:2008-09-20 Published:2008-09-20
  • Contact: XIAO Yan-Hua
  • Supported by:
     

Abstract: In order to improve the S. androgynus cold tolerance, genetic transformation system of S. androgynus was established in this study. The cDNA Mpafp149 of Microdera punctipennis dzungarica was used as target gene to obtain the cold tolerant transgenic S. androgynus plants. The stems of S. androgynus were transformed with Agrobacterium tumefaciens EHA105 strain which harbors plasmid pCAMBIA-1302-Mpafp149.The transformation parameters of Agrobacterium concentration, pre-culture period, co-culture period, immersion time and concentrations of antibiotics(hygromycin) were optimized selectively. The results showed that the better physiological and transforming condition was pre-cultured and co-cultured for 2 days respectively, immersed in OD600 0.2~0.25 liquid bacterium for 8 min. It was primarily proved by PCR that the Mpafp149 was integrated into the Sauropus androgynus and the T0 transgenic S. androgynus plants were obtained successfully.

Key words: Sauropus androgynus, insect antifreeze protein gene, agrobacterium mediated, genetic transformation

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