Biotechnology Bulletin ›› 2013, Vol. 0 ›› Issue (1): 123-128.

• Research report • Previous Articles     Next Articles

Cloning and Expression of Bloom Helicase Mutant

Zhang Jinbiao1,2,3, Xu Houqiang2,3 ,Luo Heng2,3 ,Xu Qinghe1,2,3 ,Chen Xiang2,3 ,Li Kun1,2,3   

  1. 1. College of Life Science，Guizhou University，Guiyang 550025;
    2. Key Laboratory of Animal Genetics，Breeding and Reproduction in the Plateau Mountainous Region，Ministry of Education，College of Animal Science，Guizhou University，Guiyang 550025;
    3. Guizhou Key Laboratory of Animal Genetics，Breeding Reproduction，College of Animal Science，Guizhou University，Guiyang 550025
  • Received:2012-07-06 Revised:2013-01-30 Online:2013-01-30 Published:2013-01-30

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Abstract:

Bloom syndrome(BS) which is caused by mutation of BLM is an autosomal recessive disorder characterized by genomicinstability and the high rate of many types of cancer. Disease-causing missense mutations have been identified at 1 036 cysteines localized in theRecQ-Ct domain. In this work, with bioinformatics and molecular biology technology, blm gene which 985 and 1 036 amino acids were mutated was obtained by two cycles of recombinant PCR, and cloned into expression vector pET-28a, and transformed into the strain E. coli BL21(DE3).It was purified that double mutation BLM helicase is the high purity(>95%), and identified by Western blotting. They could be pivotal forunderstanding the pathogenic mechanisms of BS.

Key words: BLM helicase , Recombinant PCR , Multisite mutation , Gene cloning and expressing