[Objective] The aim was to establish the BHK-21 stable cell lines expressing T7 RNAP and GFP.[Method]T7 RNAP gene was amplified from E.coli BL21(DE3)and inserted into FG12 vector.The lenti-virus recombinant plasmid ...[Objective] The aim was to establish the BHK-21 stable cell lines expressing T7 RNAP and GFP.[Method]T7 RNAP gene was amplified from E.coli BL21(DE3)and inserted into FG12 vector.The lenti-virus recombinant plasmid FG12-T7 RNAP plasmid was obtained via identification with double enzymes digestion and gene sequencing.The transient expressed T7 RNAP protein was determined by WB in 293T cells transfected with FG12-T7 RNAP plasmid.The recombinant FG12-RNAP lenti-virus was packaged up by transfecting the 293 cells with the recombinant vector FG12-RNAP and the helper plasmids via lipofectamine-2000,which was then used to infect BHK-21 cells.The positive cell clones were obtained after continuous screening by GFP.The expression of T7 RNAP gene was confirmed by Western blot.[Result]The cell line stably expressing the T7 RNAP gene was established by Western blot.[Conclusion]T7 RNAP gene could be stably expressed in eukaryotic cells and it provided a good platform to rescue RNA virus in vivo.展开更多
A 45-day greenhouse experiment was carried out to determine effect of vesicular-arbuscular (VA) mycorrhizal fungi on colonization rate, plant height, plant growth, hyphae length, total Al in the plants, exchangeable A...A 45-day greenhouse experiment was carried out to determine effect of vesicular-arbuscular (VA) mycorrhizal fungi on colonization rate, plant height, plant growth, hyphae length, total Al in the plants, exchangeable Al in the soil and soil pH by comparison at soil pH 3.5, 4.5 and 6.0. Plant mung bean (Phaseolus radiatus L.) and crotalaria (Crotalaria mucronata Desv.) were grown with and without VA mycorrhizal fungi in pots with red soil. Ten VA mycorrhizal fungi strains were tested, including Glomus epigaeum (No. 90001), Glomus caledonium (No. 90036), Glomus mosseae (No. 90107), Acaulospora spp. (No. 34), Scutellospora heterogama (No. 36), Scutellospora calospora (No. 37), Glomus manihotis (No. 38), Gigaspora spp. (No. 47), Glomus manihotis (No. 49), and Acaulospora spp. (No. 53). Being the most tolerant to acidity, strain 34 and strain 38 showed quicker and higher-rated colonization without lagging, three to four times more in number of nodules, two to four times more in plant dry weight, 30% to 60% more in hyphae length, lower soil exchangeable Al, and higher soil pH than without VA mycorrhizal fungi (CK). Other strains also could improve plant growth and enhance plant tolerance to acidity, but their effects were not marked. This indicated that VA mycorrhizal fungi differed in the tolerance to soil acidity and so did their inoculation effects. In the experiment, acidic soil could be remedied by inoculation of promising VA mycorrhizal fungi tolerant of acidity.展开更多
A perimeter traffic signal control strategy is proposed based on the macroscopic fundamental diagram theory(MFD)to solve the signal control problem in oversaturated states.First,the MFD of a specific regional network ...A perimeter traffic signal control strategy is proposed based on the macroscopic fundamental diagram theory(MFD)to solve the signal control problem in oversaturated states.First,the MFD of a specific regional network can be derived using VSS IM simulation software.Secondly,the maximum number of cumulative vehicles that the network can accommodate is determined based on the MFD.Then,through monitoring the influx flow,the number of vehicles existing in and exiting from the network,a perimeter traffic control model is proposed to optimize the signal timing of the boundary intersections.Finally,a virtual network simulation model is established and three different kinds o f traffic demand are loaded into the network.Simulation results show that a fer the strategy implementation,the number o f vehicles accumulating in the network can be kept near the optimal value,while the number o f both entering and exiting vehicles increases significantly and the road network can be maintained at a large capacity.Simultaneously,the queue length at the approach of the border intersections is reasonably controlled and vehicles entering and exiting the network can maintain a more efficient and stable speed.The network performance indices such as the average traffic delay and average number of stops can be improved to a certain degree,thus verifying the effectiveness and feasibility of the perimeter control strategy.展开更多
Cultural practices, hand collection of larvae, light traps and entomopathogenic fungi were investigated to manage of palm borers Oryctes spp. in date palm orchards during the years 2010-2015. Cultural practices, such ...Cultural practices, hand collection of larvae, light traps and entomopathogenic fungi were investigated to manage of palm borers Oryctes spp. in date palm orchards during the years 2010-2015. Cultural practices, such as sanitation and pruning frond bases, hand collection of larvae during regular annual service work and light traps with solar energy were practiced annually in one orchard for five years. In the second orchard, cultural practices were applied for two years, light trap for one year only, and no hand collection of larvae was done during the period. The third orchard was used as a control treatment. Results showed reduction in population density of larvae (91.6% and 53.0%) and adult (76.1% and 41.1%) ofOryects spp. in the 1st and 2nd orchard, respectively The results also demonstrated the impact of moon light phases on the number of Oryctes spp. adults caught by light trap and the existence of an inverse relation between moon light and flight activity of adults. Biological experiments also revealed that entomopathogenic fungi can cause high mortality rate reaching 100% after 29 d. Beauveria bassiana scored higher mortality rate in short time, especially at concentration of 1 × 10^11 conidia/mL with LT50 = 12.75 and LT90 = 20; while, Metarhizium anisopliae caused the higher percentage of malformed adults. The results depicted the effectiveness of some integrated ecological sound control methods for monitoring and population suppression of Oryects spp. in date palm orchards.展开更多
Wind environment simulation of a commercial district in Baise city of Guangxi Province, China, is carried out in the design phase. The results are analyzed and based on the evaluation standard for greening building of...Wind environment simulation of a commercial district in Baise city of Guangxi Province, China, is carried out in the design phase. The results are analyzed and based on the evaluation standard for greening building of China. The simulation method is discussed in detail, and some suggestions for wind environment optimization are put forward, which might be helpful for similar research.展开更多
Objective: To label human insulin-like growth factor-I (hIGF-I) eukaryotic expression vector with green fluorescent protein (GFP) for the repair of articular cartilage defects. Methods: GFP cDNA was inserted into ...Objective: To label human insulin-like growth factor-I (hIGF-I) eukaryotic expression vector with green fluorescent protein (GFP) for the repair of articular cartilage defects. Methods: GFP cDNA was inserted into pcDNA3.1-hIGF-1 to construct the co-expression vector with two multiple cloning sites mammalian expression vector under two cytomegalovirus promoters/enhancers respectively. Recombinant pcGI was transfected into NIH 3T3 cells with the help of lipofectamine. Results: Enzyme digestion and agarose gel electrophoresis analysis revealed that pcGI vector contained correct GFP and hIGF-I cDNA. Expression of (hIGF-1) and GFP was confirmed in transfected NIH 3T3 cells by immunocytochemical analysis and fluorescence microscopy. Conclusions: hIGF-I eukaryotic expression vector has been successfully labeled with GFP.展开更多
基金Supported by National Transgenic Major Program of China(2009ZX08007-006B)the National Natural Science Foundation of China(31072160)+2 种基金Science and Technique Foundation of Shandong Province(2009GG20002032)Natural Science Foundation of Shandong Province(Y2008D20)an Open Issue of State Key Laboratory of Veterinary Biotechnology Fund(SKLVBF200806)~~
文摘[Objective] The aim was to establish the BHK-21 stable cell lines expressing T7 RNAP and GFP.[Method]T7 RNAP gene was amplified from E.coli BL21(DE3)and inserted into FG12 vector.The lenti-virus recombinant plasmid FG12-T7 RNAP plasmid was obtained via identification with double enzymes digestion and gene sequencing.The transient expressed T7 RNAP protein was determined by WB in 293T cells transfected with FG12-T7 RNAP plasmid.The recombinant FG12-RNAP lenti-virus was packaged up by transfecting the 293 cells with the recombinant vector FG12-RNAP and the helper plasmids via lipofectamine-2000,which was then used to infect BHK-21 cells.The positive cell clones were obtained after continuous screening by GFP.The expression of T7 RNAP gene was confirmed by Western blot.[Result]The cell line stably expressing the T7 RNAP gene was established by Western blot.[Conclusion]T7 RNAP gene could be stably expressed in eukaryotic cells and it provided a good platform to rescue RNA virus in vivo.
基金Project (No. G1999011807) supported by the National Key Basic Research and Development Programme of China
文摘A 45-day greenhouse experiment was carried out to determine effect of vesicular-arbuscular (VA) mycorrhizal fungi on colonization rate, plant height, plant growth, hyphae length, total Al in the plants, exchangeable Al in the soil and soil pH by comparison at soil pH 3.5, 4.5 and 6.0. Plant mung bean (Phaseolus radiatus L.) and crotalaria (Crotalaria mucronata Desv.) were grown with and without VA mycorrhizal fungi in pots with red soil. Ten VA mycorrhizal fungi strains were tested, including Glomus epigaeum (No. 90001), Glomus caledonium (No. 90036), Glomus mosseae (No. 90107), Acaulospora spp. (No. 34), Scutellospora heterogama (No. 36), Scutellospora calospora (No. 37), Glomus manihotis (No. 38), Gigaspora spp. (No. 47), Glomus manihotis (No. 49), and Acaulospora spp. (No. 53). Being the most tolerant to acidity, strain 34 and strain 38 showed quicker and higher-rated colonization without lagging, three to four times more in number of nodules, two to four times more in plant dry weight, 30% to 60% more in hyphae length, lower soil exchangeable Al, and higher soil pH than without VA mycorrhizal fungi (CK). Other strains also could improve plant growth and enhance plant tolerance to acidity, but their effects were not marked. This indicated that VA mycorrhizal fungi differed in the tolerance to soil acidity and so did their inoculation effects. In the experiment, acidic soil could be remedied by inoculation of promising VA mycorrhizal fungi tolerant of acidity.
基金The National Natural Science Foundation of China(No.51308227)the Fundamental Research Funds for the Central Universities(No.201522087)+1 种基金the Science and Technology Planning Project of Guangdong Province(No.2016A030305001)the Project of Department of Communications of Guangdong Province(No.2015-02-070)
文摘A perimeter traffic signal control strategy is proposed based on the macroscopic fundamental diagram theory(MFD)to solve the signal control problem in oversaturated states.First,the MFD of a specific regional network can be derived using VSS IM simulation software.Secondly,the maximum number of cumulative vehicles that the network can accommodate is determined based on the MFD.Then,through monitoring the influx flow,the number of vehicles existing in and exiting from the network,a perimeter traffic control model is proposed to optimize the signal timing of the boundary intersections.Finally,a virtual network simulation model is established and three different kinds o f traffic demand are loaded into the network.Simulation results show that a fer the strategy implementation,the number o f vehicles accumulating in the network can be kept near the optimal value,while the number o f both entering and exiting vehicles increases significantly and the road network can be maintained at a large capacity.Simultaneously,the queue length at the approach of the border intersections is reasonably controlled and vehicles entering and exiting the network can maintain a more efficient and stable speed.The network performance indices such as the average traffic delay and average number of stops can be improved to a certain degree,thus verifying the effectiveness and feasibility of the perimeter control strategy.
文摘Cultural practices, hand collection of larvae, light traps and entomopathogenic fungi were investigated to manage of palm borers Oryctes spp. in date palm orchards during the years 2010-2015. Cultural practices, such as sanitation and pruning frond bases, hand collection of larvae during regular annual service work and light traps with solar energy were practiced annually in one orchard for five years. In the second orchard, cultural practices were applied for two years, light trap for one year only, and no hand collection of larvae was done during the period. The third orchard was used as a control treatment. Results showed reduction in population density of larvae (91.6% and 53.0%) and adult (76.1% and 41.1%) ofOryects spp. in the 1st and 2nd orchard, respectively The results also demonstrated the impact of moon light phases on the number of Oryctes spp. adults caught by light trap and the existence of an inverse relation between moon light and flight activity of adults. Biological experiments also revealed that entomopathogenic fungi can cause high mortality rate reaching 100% after 29 d. Beauveria bassiana scored higher mortality rate in short time, especially at concentration of 1 × 10^11 conidia/mL with LT50 = 12.75 and LT90 = 20; while, Metarhizium anisopliae caused the higher percentage of malformed adults. The results depicted the effectiveness of some integrated ecological sound control methods for monitoring and population suppression of Oryects spp. in date palm orchards.
文摘Wind environment simulation of a commercial district in Baise city of Guangxi Province, China, is carried out in the design phase. The results are analyzed and based on the evaluation standard for greening building of China. The simulation method is discussed in detail, and some suggestions for wind environment optimization are put forward, which might be helpful for similar research.
文摘Objective: To label human insulin-like growth factor-I (hIGF-I) eukaryotic expression vector with green fluorescent protein (GFP) for the repair of articular cartilage defects. Methods: GFP cDNA was inserted into pcDNA3.1-hIGF-1 to construct the co-expression vector with two multiple cloning sites mammalian expression vector under two cytomegalovirus promoters/enhancers respectively. Recombinant pcGI was transfected into NIH 3T3 cells with the help of lipofectamine. Results: Enzyme digestion and agarose gel electrophoresis analysis revealed that pcGI vector contained correct GFP and hIGF-I cDNA. Expression of (hIGF-1) and GFP was confirmed in transfected NIH 3T3 cells by immunocytochemical analysis and fluorescence microscopy. Conclusions: hIGF-I eukaryotic expression vector has been successfully labeled with GFP.