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Expression of the mouse metallothionein-I gene conferring cadmium resistance in a transgenic cyanobacterium
Authors:Li Ren  Dingji Shi   Jixun Dai  Binggen Ru
Affiliation:Ocean University of Qingdao, Shandong 266003, P.R. China;Lab. of Photosynthesis, Institute of Botany, Academia Sinica, Beijing 100093, P.R. China;Peking University, Peking 100871, P.R. China
Abstract:This paper reports the construction of a transgenic strain of cyanobacterium aimed at removing heavy metal pollution in waters. The mouse metallothionein-I (mMT-I) gene was inserted in the vector pRL-439 downstream of the strong psbA promoter. The resulting plasmid pRL-MT was ligated at the EcoRI site of the shuttle vector pKT-210 to generate the shuttle expression vector pKT-MT. This recombinant plasmid was introduced into Anabaena sp. PCC 7120 by triparental conjugative transfer. After selection on streptomycin, a stable transgenic Anabaena strain was obtained. The presence of the mMT-I gene was confirmed by DNA/DNA hybridization and its expression was demonstrated by immunodetection with specific antibodies. A metal tolerance experiment showed that this transgenic Anabaena strain had acquired higher metal resistance.
Keywords:Mouse metallothionein gene    Anabaena sp. PCC 7120    Transgenic cyanobacterium    Heavy metal tolerance    Triparental conjugative transfer
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