通过室内急性暴露实验研究了翡翠贻贝（Perna viridis）消化腺富集Cu及其MTLP（metallothionein like protein）水平随时间的变化规律．结果表明，2种Cu浓度暴露条件下（12．7μg／L和63．5μg／L），贻贝消化腺内Cu的平均吸收速率分别...通过室内急性暴露实验研究了翡翠贻贝（Perna viridis）消化腺富集Cu及其MTLP（metallothionein like protein）水平随时间的变化规律．结果表明，2种Cu浓度暴露条件下（12．7μg／L和63．5μg／L），贻贝消化腺内Cu的平均吸收速率分别为2．045和7．028μg·（g·d）^-1，富集系数分别为2074和1619．实验测定了所有样本的溶解态Cu与总Cu含量，2个暴露组二者的比值随时间呈现出不同的变化趋势，低浓度组先降低随后上升到与对照样本几乎相同的水平，而高浓度组一直呈下降趋势，表明不同污染程度生物体内金属消化机制进程存在差异．利用Brdicka极谱法测定了贻贝消化腺内MTLP的含量，对照组贻贝消化腺的MTLP平均含量为（0．551±0．037）ms／g；12．7μg／L Cu暴露组MTLP含量随时间的变化范围是0．407～0．699ms／g，而63．5μg／L Cu暴露组在暴露初始MTLP水平就显著增加（P〈0．001），变化范围由初始0．942mg／g降至0．826mg／g．分析结果表明贻贝消化腺内的MTLP水平随着水体及生物体内的金属含量升高而增加，并与体内Cu浓度成明显的负指数增长关系（P〈0．0001）．展开更多
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. reinh...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.展开更多
This paper studies the rapid identification of heavy metal-binding peptides (phytochelatin) by taking Water Hyacinth as a model plant. Plants were cultured in water containing 2 μg/ml Cd2+ for 13 days. The Sephadex G...This paper studies the rapid identification of heavy metal-binding peptides (phytochelatin) by taking Water Hyacinth as a model plant. Plants were cultured in water containing 2 μg/ml Cd2+ for 13 days. The Sephadex G-50 chromatography of root extract under low salt concentration (0. 01 mol/L PBS) gave a Cd-binding peak with MW of 10 ,000 determined by SEC HPLC. After oxidation with performic acid, its SEC HPLC molecular weight decreased to below 1300 and the reverse phase HPLC showed one peptide peak, whose amino acid composition is the same as that of the sample never undertaking oxidation, and (Glu/Gln):Cys:Gly=2:2:1. According to the general structure of phytochelatin (γ-Glu-Cys)n-Gly, n is 2 in this case. The protocol including the sequential steps of Sephadex G-50 chromatography→performic acid oxidation→reverse phase HPLC→amino acid analysis is a rapid and effective method to identify the existence of phytochelatin and determine its values of n.展开更多
基金Supported by the Natural Science Foundation of Guangdong Province (Grant No. 5010492)the Plan of Science and Technology of Shenzhen Municipality, China
文摘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.
文摘This paper studies the rapid identification of heavy metal-binding peptides (phytochelatin) by taking Water Hyacinth as a model plant. Plants were cultured in water containing 2 μg/ml Cd2+ for 13 days. The Sephadex G-50 chromatography of root extract under low salt concentration (0. 01 mol/L PBS) gave a Cd-binding peak with MW of 10 ,000 determined by SEC HPLC. After oxidation with performic acid, its SEC HPLC molecular weight decreased to below 1300 and the reverse phase HPLC showed one peptide peak, whose amino acid composition is the same as that of the sample never undertaking oxidation, and (Glu/Gln):Cys:Gly=2:2:1. According to the general structure of phytochelatin (γ-Glu-Cys)n-Gly, n is 2 in this case. The protocol including the sequential steps of Sephadex G-50 chromatography→performic acid oxidation→reverse phase HPLC→amino acid analysis is a rapid and effective method to identify the existence of phytochelatin and determine its values of n.
