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Journal of Shanghai University（Natural Science Edition）

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Construction and Characterization of a Dunaliella viridis Bacterial Artificial Chromosome Library

MENG Xiang zong;YANG Xiao wu;GUAN Zhen wei;et al   

  1. 1. School of Life Sciences, Shanghai University, Shanghai 200444, China; 2. Shanghai Key Laboratory of Bio energy Crop, Shanghai 200444, China; 3. Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Scienc es, Shanghai 200032, China
  • Received:2006-04-06 Online:2007-02-28 Published:2007-02-28

Abstract: Dunaliella viridis has outstanding ability to adapt to hyper salinity, and is a model system for plant salt tolerance mechanism study. We constructed a bac terial artificial chromosome library of Dunaliella viridis , using vector pCC 1 BAC. The library consisted of 9 216 clones on 96 96 well plates. The average in sertion size was about 55 kb. A four dimensional PCR screening system was devel oped, by which single positive clone could be obtained with 4 rounds of PCR. Bas ed on the sequences of two cDNAs of Dunaliella viridis isolated in our lab ( DvSPT2 and DvTPSP respectively), we isolated 4 positive clones for each gene by PCR, indicating that the library had about 4 times genome coverage.

Key words: bacterial artificial chromosome (BAC) library, four dim ensional PCR (4D PCR) screening system, Dunaliella viridis