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Journal of Shanghai University（Natural Science Edition） ›› 2011, Vol. 17 ›› Issue (5): 675-680.doi: 10.3969/j.issn.1007-2861.2011.05.019

• Life Science • Previous Articles     Next Articles

Construction and Characterization of Bacterial Artificial Chromosome Library of Cupriavidus necator JMP134

ZHANG Jun-ya,SONG Ren-tao,XU Zheng-kai,ZHU Chen-guang   

  1. Shanghai Key Laboratory of Bio-energy Crops, School of Life Sciences, Shanghai University, Shanghai 200444, China
  • Received:2010-03-31 Online:2011-10-26 Published:2011-10-26

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Abstract: Cupriavidus necator JMP134 (C.necator JMP134) is a model for biodegradation of aromatic compounds. The function of phenol degradation of this strain relies on a whole gene cluster. Therefore, isolation of the gene cluster is important. A bacterial artificial chromosome (BAC) library from C. necator JMP134 with vector pIndigo-BAC 5 was constructed. The library consisted of about 9 600 clones, 99% of which having an insertion with average insertion size of 65 kb. The BAC library was estimated to have 860 times of genome size. After screening one tenth of the total library clones with PCR using the designed specific primers, eleven phlN2 positive clones were obtained. The result indicates that the BAC library has high quality and high coverage, and is sufficient for isolation of target genes.

Key words: Cupriavidus necator JMP134, bacterial artificial chromosome (BAC) library, phenol biodegradation

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