Expression and function analysis of the metallo-thionein-like (MT-like) gene from Festuca rubra in Chlamydomonas reinhardtii chloroplast  

作　　者：HAN SiHai1,2, HU ZhangLi1 & LEI AnPing1 1 Institute of Eco-Environment, College of Life Science, Shenzhen University, Shenzhen 518060, China 2 Research Center for Analytical Instrumentation, State Key Lab of Industrial Control Technology, Institute of Cyber-Systems and Control, Zhejiang University, Hangzhou 310058, China 

出　　处：《Science China(Life Sciences)》2008年第12期1076-1081,共6页中国科学（生命科学英文版）

基　　金：Supported by the Natural Science Foundation of Guangdong Province (Grant No. 5010492);the Plan of Science and Technology of Shenzhen Municipality, China

摘　　要：The cDNA of the metallothionein-like (MT-like) gene from Festuca rubra cv. Merlin was optimized with bias codon of Chlamydomonous reinhardtii chloroplast genome. The optimized MT-like gene was de-livered into C. reinhardtii chloroplast and the transgenic strains expressing MT-like gene was obtained. PCR-Southern blot and RT-PCR-Southern blot analysis demonstrated that the MT-like gene was inte-grated into chloroplast genome of C. reinhardtii and expressed at the transcriptional level. The cad-mium binding capacity of the transgenic C. reinhardtii was determined by hydride generation-atomic fluorescence spectrometry (HG-AFS) and the binding properties were analyzed. Results showed that the transgenic C. reinhardtii expressing the MT-like gene exhibited remarkably higher Cd2+ binding capacity and grew to higher densities at toxic Cd2+ concentrations (40-100 μmol/L) than the wild type strain, and that the IC50 of Cd2+ (3-d treating ) to algal cell growth of transgenic strain was 55.43% higher than that of the wild type strain, indicating that the Cd2+ binding capacity and Cd2+ tolerance of C. reinhardtii was enhanced through the expression of the foreign MT-like gene in chloroplast.The cDNA of the metallothionein-like (MT-like) gene from Festuca rubra cv. Merlin was optimized with bias codon of Chlamydomonous reinhardtii chloroplast genome. The optimized MT-like gene was de-livered into C. reinhardtii chloroplast and the transgenic strains expressing MT-like gene was obtained. PCR-Southern blot and RT-PCR-Southern blot analysis demonstrated that the MT-like gene was inte-grated into chloroplast genome of C. reinhardtii and expressed at the transcriptional level. The cad-mium binding capacity of the transgenic C. reinhardtii was determined by hydride generation-atomic fluorescence spectrometry (HG-AFS) and the binding properties were analyzed. Results showed that the transgenic C. reinhardtii expressing the MT-like gene exhibited remarkably higher Cd2+ binding capacity and grew to higher densities at toxic Cd2+ concentrations (40-100 μmol/L) than the wild type strain, and that the IC50 of Cd2+ (3-d treating ) to algal cell growth of transgenic strain was 55.43% higher than that of the wild type strain, indicating that the Cd2+ binding capacity and Cd2+ tolerance of C. reinhardtii was enhanced through the expression of the foreign MT-like gene in chloroplast.

关 键 词：CHLAMYDOMONAS REINHARDTII CHLOROPLAST metallothionein-like (MT-like) GENE cadmium binding properties 

分 类 号：Q943.2[生物学—植物学]