The agp gene encoding the ADP-glucose pyrophosphorylase involved in cyanobacterial glycogen synthesis was amplified by PCR. The resulting agp fragment was cloned in plasmid pUC118 to generate plasmid pUCA. Part of the...The agp gene encoding the ADP-glucose pyrophosphorylase involved in cyanobacterial glycogen synthesis was amplified by PCR. The resulting agp fragment was cloned in plasmid pUC118 to generate plasmid pUCA. Part of the fragment within the agp DNA was deleted and replaced by an erythromycin resistance cassette to generate plasmid pUCAE, which was used to transform the Synechocystis sp. PCC 6803 wild-type strain and a mutant with resistance to erythromycin was obtained. PCR analysis of the genomic DNA from the resulting mutant indicated that the appropriate deletion and insertion indeed had occurred. The cell growth and Chl a, glycogen content in the mutant showed difference from those in the wild-type strain. The obtained biomass as well as the Chl a content in the mutant strain was higher than that of the wild-type strain, which suggested that the photosynthesis efficiency in the agp(-) strain was higher than that in the wild-type strain. No glycogen was found in the mutant, providing evidence for the correction of the mutant in physiological level.展开更多
Genetic modifi cation is useful for improving the nutritional qualities of cyanobacteria. To increase the total unsaturated fatty acid content, along with the ratio of ω-3/ω-6 fatty acids, genetic engineering can be...Genetic modifi cation is useful for improving the nutritional qualities of cyanobacteria. To increase the total unsaturated fatty acid content, along with the ratio of ω-3/ω-6 fatty acids, genetic engineering can be used to modify fatty acid metabolism. S ynechococcus sp. PCC7002, a fast-growing cyanobacterium, does not contain a Δ6 desaturase gene and is therefore unable to synthesize γ-linolenic acid(GLA) and stearidonic acid(SDA), which are important in human health. In this work, we constructed recombinant vectors Syd6 D, Syd15 D and Syd6Dd15 D to express the Δ15 desaturase and Δ6 desaturase genes from Synechocystis PCC6803 in Synechococcus sp. PCC7002, with the aim of expressing polyunsaturated fatty acids. Overexpression of the Δ15 desaturase gene in S ynechococcus resulted in 5.4 times greater accumulation of α-linolenic acid compared with the wild-type while Δ6 desaturase gene expression produced both GLA and SDA. Co-expression of the two genes resulted in low-level accumulation of GLA but much larger amounts of SDA, accounting for as much to 11.64% of the total fatty acid content.展开更多
The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the ...The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the present study. The species was cultured in BG11 liquid medium contained various initial concentrations of Pb^2＋ and Cd〉 （0, 0.5, 1, 2, 4, 6 and 8 mg/L）. The experiment was conducted for six days and the metal induced alterations in the ultrastructure, growth and pigment contents were assessed. Alterations in the ultrastructure of the Synechocystis sp. PCC 6803 cells became evident with the increased （〉4 mg/L Pb^2＋） metal concentration. The photosynthetic apparatus （thylakoid membranes） were found to be the worst affected. Deteriorated or completely destroyed thylakoid membranes have made large empty spaces in the cell interior. In addition, at the highest concentration （8 mg/L pb^2＋）, the polyphosphate granules became more prominent both in size and number. Despite the initial slight stimulations （0.2, 3.8 and 6.5% respectively at 0.5, 1 and 2 mg/L pb^2＋）, both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents （chlorophyll a, 13 carotene and phycocyanin） were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb^2＋, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll a, 13 carotene and phycocyanin respectively over the control. Corresponding reductions for the same CdZ＋concentrations were 57.83, 48.94 and 56.90%. Lethal concentration （96 h LC50） values （3.47 mg/L Cd^2＋ and 12.11 mg/L Pb^2＋） indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd^2＋ than Pb^2＋.展开更多
Based on analyzing the overlap appearance of random winding, basic principles of ribbon-free random winding and parametric selections for anti-overlap are discussed. The ribbon-free random winding control system and i...Based on analyzing the overlap appearance of random winding, basic principles of ribbon-free random winding and parametric selections for anti-overlap are discussed. The ribbon-free random winding control system and its related hardware construction as well as its software design are introduced by applying PCC technology to high speed spinning machine of synthetic filament.展开更多
ThefecC gene encoding a putative iron (III) dicitrate transporter was cloned from nitrogen-fixing cyanobacteriumAnabaena sp. PCC 7120, and inactivated. The mutant grows normally in medium with NO 3 ? , NH 4 + or witho...ThefecC gene encoding a putative iron (III) dicitrate transporter was cloned from nitrogen-fixing cyanobacteriumAnabaena sp. PCC 7120, and inactivated. The mutant grows normally in medium with NO 3 ? , NH 4 + or without combined nitrogen. But in iron-deficient medium, the mutant grows slowly. Photosynthetic properties were compared between the mutant and the wildtype strain, the content of photosynthetic pigments in the mutant is lower than that of the wild-type. The results of RT-PCR experiments show that thefecC gene is expressed under iron-deficient conditions, but is not expressed under iron-replete conditions. These results revealed thatfecC gene product is required for optimal growth under iron-deficient conditions inAnabaena sp. PCC 7120. Key words Anabaena sp. PCC 7120 - fecC - iron deficiency - photosynthetic properties - expression CLC number Q 933 Foundation item: Supported by the National Natural Sciences Foundation of China (30070154), the Frontier Science Projects Program of the Institute of Hydrobiology, the Chinese Academy of Sciences (220316), State Key Project on Cyanobacterial Bloom Control in Lake Dianchi (K99-05-35-01)Biography: XU Wen-liang (1974-), male, Ph. D, research direction: molecular genetics of cyanobacteria.展开更多
Inspired by iron fertilization experiments in HNLC（high-nitrate, low-chlorophyll） sea areas,we proposed the use of iron-rich engineered microalgae for microbial contaminant control in iron-free culture media. Based ...Inspired by iron fertilization experiments in HNLC（high-nitrate, low-chlorophyll） sea areas,we proposed the use of iron-rich engineered microalgae for microbial contaminant control in iron-free culture media. Based on the genome sequence and natural transformation system of Synechocystis sp. PCC6803, ftn A（encoding ferritin） was selected as our target gene and was cloned into wild-type Synechocystis sp. PCC6803. Tests at the molecular level confirmed the successful construction of the engineered Synechocystis sp. PCC6803-ftn A. After Fe3＋-EDTA pulsing, the intracellular iron content of Synechocystis sp. PCC6803-ftn A was significantly enhanced, and the algae was used in the microbial contamination control system. In the coupled Synechocystis sp. PCC6803-ftn A production and municipal wastewater（MW, including Scenedesmus obliquus and Bacillus） treatment, Synechocystis sp. PCC6803-ftn A accounted for all of the microbial activity and significantly increased from 70% of the microbial community to 95%.These results revealed that while the stored iron in the Synechocystis sp. PCC6803-ftn A cells was used for growth and reproduction of this microalga in the MW, the growth of other microbes was inhibited because of the iron limitation, and these results provide a new method for microbial contamination control during a coupling process.展开更多
随着移动通信业务的迅速发展,而资源的精准投放就成为运营商亟待解决的问题和关注的重点。为了降低运营成本,同时提升服务质量和用户满意度,以5G用户的营销推广为例,研究从用户筛选到营销全流程服务体系。基于机器学习模型和大数据分析...随着移动通信业务的迅速发展,而资源的精准投放就成为运营商亟待解决的问题和关注的重点。为了降低运营成本,同时提升服务质量和用户满意度,以5G用户的营销推广为例,研究从用户筛选到营销全流程服务体系。基于机器学习模型和大数据分析,利用信令数据,建立终端筛选模型及潜在用户预测模型。并通过策略控制和计费(PCC,Policy Control and Charging)匹配用户行为触点推送信息,达到精准营销减少投诉的优化体系。展开更多
文摘The agp gene encoding the ADP-glucose pyrophosphorylase involved in cyanobacterial glycogen synthesis was amplified by PCR. The resulting agp fragment was cloned in plasmid pUC118 to generate plasmid pUCA. Part of the fragment within the agp DNA was deleted and replaced by an erythromycin resistance cassette to generate plasmid pUCAE, which was used to transform the Synechocystis sp. PCC 6803 wild-type strain and a mutant with resistance to erythromycin was obtained. PCR analysis of the genomic DNA from the resulting mutant indicated that the appropriate deletion and insertion indeed had occurred. The cell growth and Chl a, glycogen content in the mutant showed difference from those in the wild-type strain. The obtained biomass as well as the Chl a content in the mutant strain was higher than that of the wild-type strain, which suggested that the photosynthesis efficiency in the agp(-) strain was higher than that in the wild-type strain. No glycogen was found in the mutant, providing evidence for the correction of the mutant in physiological level.
基金Supported by the International S&T Cooperation Program of China(No.2012DFA30450)the National Natural Science Foundation of China(No.30871541)+1 种基金the Taishan Scholar Foundation of Shandong Province(No.tshw20091014)the Innovation Program of the University Institutes of Jinan,Shandong Province(No.201004044)
文摘Genetic modifi cation is useful for improving the nutritional qualities of cyanobacteria. To increase the total unsaturated fatty acid content, along with the ratio of ω-3/ω-6 fatty acids, genetic engineering can be used to modify fatty acid metabolism. S ynechococcus sp. PCC7002, a fast-growing cyanobacterium, does not contain a Δ6 desaturase gene and is therefore unable to synthesize γ-linolenic acid(GLA) and stearidonic acid(SDA), which are important in human health. In this work, we constructed recombinant vectors Syd6 D, Syd15 D and Syd6Dd15 D to express the Δ15 desaturase and Δ6 desaturase genes from Synechocystis PCC6803 in Synechococcus sp. PCC7002, with the aim of expressing polyunsaturated fatty acids. Overexpression of the Δ15 desaturase gene in S ynechococcus resulted in 5.4 times greater accumulation of α-linolenic acid compared with the wild-type while Δ6 desaturase gene expression produced both GLA and SDA. Co-expression of the two genes resulted in low-level accumulation of GLA but much larger amounts of SDA, accounting for as much to 11.64% of the total fatty acid content.
文摘The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the present study. The species was cultured in BG11 liquid medium contained various initial concentrations of Pb^2＋ and Cd〉 （0, 0.5, 1, 2, 4, 6 and 8 mg/L）. The experiment was conducted for six days and the metal induced alterations in the ultrastructure, growth and pigment contents were assessed. Alterations in the ultrastructure of the Synechocystis sp. PCC 6803 cells became evident with the increased （〉4 mg/L Pb^2＋） metal concentration. The photosynthetic apparatus （thylakoid membranes） were found to be the worst affected. Deteriorated or completely destroyed thylakoid membranes have made large empty spaces in the cell interior. In addition, at the highest concentration （8 mg/L pb^2＋）, the polyphosphate granules became more prominent both in size and number. Despite the initial slight stimulations （0.2, 3.8 and 6.5% respectively at 0.5, 1 and 2 mg/L pb^2＋）, both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents （chlorophyll a, 13 carotene and phycocyanin） were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb^2＋, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll a, 13 carotene and phycocyanin respectively over the control. Corresponding reductions for the same CdZ＋concentrations were 57.83, 48.94 and 56.90%. Lethal concentration （96 h LC50） values （3.47 mg/L Cd^2＋ and 12.11 mg/L Pb^2＋） indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd^2＋ than Pb^2＋.
文摘Based on analyzing the overlap appearance of random winding, basic principles of ribbon-free random winding and parametric selections for anti-overlap are discussed. The ribbon-free random winding control system and its related hardware construction as well as its software design are introduced by applying PCC technology to high speed spinning machine of synthetic filament.
文摘ThefecC gene encoding a putative iron (III) dicitrate transporter was cloned from nitrogen-fixing cyanobacteriumAnabaena sp. PCC 7120, and inactivated. The mutant grows normally in medium with NO 3 ? , NH 4 + or without combined nitrogen. But in iron-deficient medium, the mutant grows slowly. Photosynthetic properties were compared between the mutant and the wildtype strain, the content of photosynthetic pigments in the mutant is lower than that of the wild-type. The results of RT-PCR experiments show that thefecC gene is expressed under iron-deficient conditions, but is not expressed under iron-replete conditions. These results revealed thatfecC gene product is required for optimal growth under iron-deficient conditions inAnabaena sp. PCC 7120. Key words Anabaena sp. PCC 7120 - fecC - iron deficiency - photosynthetic properties - expression CLC number Q 933 Foundation item: Supported by the National Natural Sciences Foundation of China (30070154), the Frontier Science Projects Program of the Institute of Hydrobiology, the Chinese Academy of Sciences (220316), State Key Project on Cyanobacterial Bloom Control in Lake Dianchi (K99-05-35-01)Biography: XU Wen-liang (1974-), male, Ph. D, research direction: molecular genetics of cyanobacteria.
基金supported by the National Key Technologies R&D Program of China(No.2012BAJ25B02)
文摘Inspired by iron fertilization experiments in HNLC（high-nitrate, low-chlorophyll） sea areas,we proposed the use of iron-rich engineered microalgae for microbial contaminant control in iron-free culture media. Based on the genome sequence and natural transformation system of Synechocystis sp. PCC6803, ftn A（encoding ferritin） was selected as our target gene and was cloned into wild-type Synechocystis sp. PCC6803. Tests at the molecular level confirmed the successful construction of the engineered Synechocystis sp. PCC6803-ftn A. After Fe3＋-EDTA pulsing, the intracellular iron content of Synechocystis sp. PCC6803-ftn A was significantly enhanced, and the algae was used in the microbial contamination control system. In the coupled Synechocystis sp. PCC6803-ftn A production and municipal wastewater（MW, including Scenedesmus obliquus and Bacillus） treatment, Synechocystis sp. PCC6803-ftn A accounted for all of the microbial activity and significantly increased from 70% of the microbial community to 95%.These results revealed that while the stored iron in the Synechocystis sp. PCC6803-ftn A cells was used for growth and reproduction of this microalga in the MW, the growth of other microbes was inhibited because of the iron limitation, and these results provide a new method for microbial contamination control during a coupling process.
文摘随着移动通信业务的迅速发展,而资源的精准投放就成为运营商亟待解决的问题和关注的重点。为了降低运营成本,同时提升服务质量和用户满意度,以5G用户的营销推广为例,研究从用户筛选到营销全流程服务体系。基于机器学习模型和大数据分析,利用信令数据,建立终端筛选模型及潜在用户预测模型。并通过策略控制和计费(PCC,Policy Control and Charging)匹配用户行为触点推送信息,达到精准营销减少投诉的优化体系。
