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Expression of Mammalian Metallothionein-I Gene in Cyanobacteria to Enhance Heavy Metal Resistance |
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| Authors: | ZChen L Ren Q Shao D Shi and B Ru |
| Institution: | a College of Life Science, Peking University, Beijing, People's Republic of China b Institute of Botany, Chinese Academy of Science, Beijing, People's Republic of China |
| Abstract: | Metallothionein (MT) from mammals is a low molecular weight (about 6–7 KD), and cystein-rich (20 cystein per molecular) protein (Vallee, 1991). So MT has the strong binding ability of metal such as Cd, Hg and Pd. Wild-typed cyanobacteria have their own metallothionein-like proteins, which are of low cysteine content and have weak ability to bind heavy metal. Therefore, it is imperative to transfer the MT-I gene from mammal into cyanobacteria cell for the absorption and accumulation of heavy metals in aqueous system including marine and lake ( Misra and Gedamu, 1989). We inserted the MT-I cDNA gene after the strong promoter PpsbA or Psmt into the intermediary vector pRL-439, and then pRL-MT was ligated with shuttle vector pKT-210 (a shuttle vector pDC-08 was used at an earlier time) to construct the shuttle expression vector pKT-MT. (The MT-I mutant  -KKS- also was selected as aim gene.) Then the constructed vector was introduced into cyanobacterium Anabaena sp. PCC7120, Synechocystis sp. PCC6803 and Synechococcus sp. PCC7942 with triparental conjunctive transfer (Bryant, 1994). The expression efficiency is about 1%. We also used homologous recombination vector pTZ18-8 to introduce MT-I cDNA into chromosomes in cyanobacteria and got higher expression efficiency. Metal tolerance tests show that the transgenic cyanobacteria acquire metal resistance up to 60 μmol/g CdCl2 and expression efficiency up to 1045 μg MT/g fresh cells according to the data of ELISA. |
| Keywords: | metallothionein metallothionein mutant gene transgenic cyanobacterium aqueous pollution heavy metal tolerance triparental conjugate transfer homologous recombination |
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