Rxol cloned from maize is a non-host gene resistant to bacterial leaf streak of rice. pCAMBIA1305-1 with Rxo1 was digested with Sca I and NgoM IV and the double right-border binary vector pMNDRBBin6 was digested with ...Rxol cloned from maize is a non-host gene resistant to bacterial leaf streak of rice. pCAMBIA1305-1 with Rxo1 was digested with Sca I and NgoM IV and the double right-border binary vector pMNDRBBin6 was digested with Hpa I and Xma I. pMNDRBBin6 carrying the gene Rxo1 was acquired by ligation of blunt-end and cohesive end. The results of PCR, restriction enzyme analysis and sequencing indicated that the Rxo1 gene had been cloned into pMNDRBBin6. This double right-border binary vector, named as pMNDRBBin6-Rxol, will play a role in breeding marker-free plants resistant to bacterial leaf streak of rice by genetic transformation.展开更多
[Objective] This study aimed to construct Brassica napus chloroplast multi- cistron double cross-over expression vector, to lay the foundation for the genetic engi- neering research of Brassica napus chloroplast. [Met...[Objective] This study aimed to construct Brassica napus chloroplast multi- cistron double cross-over expression vector, to lay the foundation for the genetic engi- neering research of Brassica napus chloroplast. [Method] Two primers were designed based on the known Brassica napus chloroplast DNA sequences AF267640 and Z50868 in GenBank. By using PCR method, two Brassica napus L. chloroplast DNA fragments were obtained, which were named RbcL and ACCD. The two Brassica na- pus chloroplast DNA homologous fragments were then cloned into plasmid pMD18-T to obtain recombinant plasmid pHBM715. Tandem expression cassette harboring spectinomycin-resistant gene aadA, mannanase gene man and green fluorescent pro- tein gene gfp was cloned into the plasmid pHBM715, thereby constructing Brassica napus chloroplast multicistron double cross-over expression vector pHBM716, which was transformed into Escherichia coil for expression and identification. [Result] Plate qualitative analysis was conducted for the functional identification of expression cas- sette in the constructed Brassica napus chloroplast multicistron double cross-over ex- pression vector, results showed that the three genes of the same multicistron were all expressed in E. coil [Conclusion] This study successfully constructed Brassica napus chloroplast multicistron double cross-over expression vector, which laid the foundation for the genetic engineering of Brassica napus chloroplast.展开更多
We have developed a 'double T-DNA' binary vector system for generating selectable marker-free transgenic plants by Agrobacterium-mediated transformation. The 'double T-DNA' binary vector pDLBRBbarm whi...We have developed a 'double T-DNA' binary vector system for generating selectable marker-free transgenic plants by Agrobacterium-mediated transformation. The 'double T-DNA' binary vector pDLBRBbarm which carried two independent T-DNAs, one containing a selectable marker neomycin phosphotransferase (nptII) gene and the other a bargene, was constructed. Transgenic tobacco (Nicotiana tabacum L.) plants were then produced by Agrobacterium-mediated transformation with this vector. Frequency of the primary transformants co-integrated with npt II gene and bar gene was 59.2%. Segregation of two T-DNA regions was found in 3 out of 4 T-1 lines from co-transformed T-0 plants with nptII and bar PPT-resistant and kanamycin-sensitive plants were in approximate 19.5% of the T-1 plants. The result indicated that this 'double T-DNA' vector system could be a workable approach to generate transgenic plants free from selectable marker genes. Co-transformation of nptII gene and bar gene to plants with mixtures of Agrobacterium tumefaciens strains containing single T-DNA vectors was also tested. Frequency of co-transformed plants was 20.0%-47.7% and relatively low as compared with that of 'double T-DNA' vector system.展开更多
In order to overcome the interference of the internal promoter in retroviral vector with a foreign DNA expression, we used the double-copy vector(DC vector) system to improve the effect of transduced gene expression.H...In order to overcome the interference of the internal promoter in retroviral vector with a foreign DNA expression, we used the double-copy vector(DC vector) system to improve the effect of transduced gene expression.Human G-CSF cDNA was inserted into the Bgl Ⅱ site of the polyclonal sites within the U3 region of the 3'long terminal repeat(3'-LTR) in the vector(N2A). After being identified, the gene was transduced into Ψ-2 packaging cell line by using the electroporation method. Consequently, the gene was duplicated in the infected cells, and transferred to the 5'-LRT, and then placed outside the retroviral transcriptional unit. After two weeks the neomycin resistance positive colonies were grown in the G-418 medium. The supernatant virus was titred and then transferred to NIH3T3 cells. The G-CSF value of positive clone N2AG7-4 was up to 53. 3 U/106 cells. Southern blot and Northern blot analysis showed that the chimeric gene faithfully duplicated in the cells infected with the corresponding virus and generated two copies with one in each LTR.展开更多
Copy-paste forgery is a very common type of forgery in JPEG images.The tampered patch has always suffered from JPEG compression twice with inconsistent block segmentation.This phenomenon in JPEG image forgeries is cal...Copy-paste forgery is a very common type of forgery in JPEG images.The tampered patch has always suffered from JPEG compression twice with inconsistent block segmentation.This phenomenon in JPEG image forgeries is called the shifted double JPEG(SDJPEG) compression.Detection of SDJPEG compressed image patches can make crucial contribution to detect and locate the tampered region.However,the existing SDJPEG compression tampering detection methods cannot achieve satisfactory results especially when the tampered region is small.In this paper,an effective SDJPEG compression tampering detection method utilizing both intra-block and inter-block correlations is proposed.Statistical artifacts are left by the SDJPEG compression among the magnitudes of JPEG quantized discrete cosine transform(DCT) coefficients.Firstly,difference 2D arrays,which describe the differences between the magnitudes of neighboring JPEG quantized DCT coefficients on the intrablock and inter-block,are used to enhance the SDJPEG compression artifacts.Then,the thresholding technique is used to deal with these difference 2D arrays for reducing computational cost.After that,co-occurrence matrix is used to model these difference 2D arrays so as to take advantage of second-order statistics.All elements of these co-occurrence matrices are served as features for SDJPEG compression tampering detection.Finally,support vector machine(SVM) classifier is employed to distinguish the SDJPEG compressed image patches from the single JPEG compressed image patches using the developed feature set.Experimental results demonstrate the efficiency of the proposed method.展开更多
A technique based on the double Fourier series is developed to estimate the winds at different isobaric levels forthe limited area domain, 35°E to 140°E and 30°S to 40°N, using the observed winds a...A technique based on the double Fourier series is developed to estimate the winds at different isobaric levels forthe limited area domain, 35°E to 140°E and 30°S to 40°N, using the observed winds at 850 hPa lcvcl for the month ofJune. For this purpose the wind field at a level under consideration is taken in the ratio form with that of 850 hPa level and the coefficients of the double Fouricr series are computed. These coefficients are subsequently used to computethe winds which are compared with the actual winds. The results of the double Fourier series technique are comparedwith those of the polynomial surface fitting method developed by Bavadekar and Khaladkar (1 992). The technique isalso applied for the daily wind data of 11. June, 1979 and the validation of the technique is tested for a few radiosondestations of india. The computed winds for these radiosonde stations arc quite close to observed winds.展开更多
DFSM （doubly fed synchronous machine） presents several advantages such as efficiency improvement, weight reduction and increase of the utilization factor （kW/kg）. In this paper the authors focus on impact of the D...DFSM （doubly fed synchronous machine） presents several advantages such as efficiency improvement, weight reduction and increase of the utilization factor （kW/kg）. In this paper the authors focus on impact of the DFSM on the efficiency and machine weight in comparison to conventional synchronous generator with wound rotor. Different topologies of DFSM are briefly described and the different methods and models for performances prediction are presented.展开更多
A modeling method of the support vector machine combined with matrix optics is considered; a complete new measurement model for double-four quadrant photoelectric detector is built. According to the analysis of the re...A modeling method of the support vector machine combined with matrix optics is considered; a complete new measurement model for double-four quadrant photoelectric detector is built. According to the analysis of the received light spot size and its motion with the changes of the defocusing amount of detector photosensitive surface and the detector position attitude in the optical path, a mathematic expression of photoelectrical conversion is given, which can be applicable to random setting position of the detector at any time. Based on least square support vector machine (LS SVM), the mapping relationship among the output signal linear characteristic parameters (zero neighborhood gradient and intercept), the defocusing amount of the detector and the installation position attitude angle is established. Thus, the multiple dimensional high accuracy measuring and adjusting control system can be left out, and adaptive measurement of the detector parameters can be realized. Compared with existed measurement model and method, the presented model has the advantages of more clear physical meaning, closer to work mechanism of detector, acquiring more complete sample data and wiping out the dead spots or bad spots in measurement. And the accuracy of displacement measurement is increased to 3?μm. At the same time, this measurement mode provides a technical shortcut for three-dimensional small angle measurement.展开更多
Expression vector pE14 with double resistance to virus and insect was constructed by inserting CW-cp gene and Bt-toxin gene one by into T-DNA of the same binary vector pE3. Tobacco was then transformed with Agrbacter...Expression vector pE14 with double resistance to virus and insect was constructed by inserting CW-cp gene and Bt-toxin gene one by into T-DNA of the same binary vector pE3. Tobacco was then transformed with Agrbacterium tumefaciens (At)GV311-SE carrying PE14. Nopaline assay and PCR amplification confirmed that both CW-cp gene and Bt-toxin gene had been introduced into tobacco genome by T-DNA of PE3. Test attack with virus and demonstrated, in some of the transgenic plants, double resistance to both infection by CMV and damage by Manduca sexta.展开更多
为了解决多功能视频编码(versatile video coding,VVC)标准下具有相同编码参数的视频双压缩检测方法准确率不高的问题,提出了一种基于编码单元(coding unit,CU)尺寸、划分模式和预测模式的检测方法。对待检测的视频进行多次编解码,分析...为了解决多功能视频编码(versatile video coding,VVC)标准下具有相同编码参数的视频双压缩检测方法准确率不高的问题,提出了一种基于编码单元(coding unit,CU)尺寸、划分模式和预测模式的检测方法。对待检测的视频进行多次编解码,分析并确定VVC流中与压缩编码次数密切相关的基础码流特征;以CU尺寸、划分模式和预测模式构建高级码流特征输入支持向量机完成视频的双压缩检测。实验结果表明,与对比文献的方法相比,所提方法的视频双压缩检测准确率有较大提升,平均准确率达到了95.82%。展开更多
针对永磁同步电机下桥臂电流采样拓扑的无位置传感器控制系统,在低载波运行时受数字延时影响易导致位置估计精度低、电流纹波增大、稳定性差等问题,提出一种半周期计算、单采样双更新的空间矢量脉宽调制方法,有效减少系统数字延时。该...针对永磁同步电机下桥臂电流采样拓扑的无位置传感器控制系统,在低载波运行时受数字延时影响易导致位置估计精度低、电流纹波增大、稳定性差等问题,提出一种半周期计算、单采样双更新的空间矢量脉宽调制方法,有效减少系统数字延时。该方法在每个脉冲宽度调制(Pulse-Width Modulation,PWM)的载波过零处采样一次电流,并在采样结束后的半周期内完成目标电压矢量计算。考虑数字延时中电机转过的角度,将目标电压矢量在下一个PWM周期的前、后半周期输出,并分别补偿0.75 T s和1.25 T s等效延时(T s为PWM周期时间),以减小目标电压矢量与实际输出电压矢量之间的误差。将新方法应用于采用模型参考自适应速度观测器的永磁同步电机无位置传感器控制系统中,实验结果表明该方法位置估计精度高、电流纹波小,且可以稳定运行于额定负载、载波比为4.0的工况。展开更多
文摘Rxol cloned from maize is a non-host gene resistant to bacterial leaf streak of rice. pCAMBIA1305-1 with Rxo1 was digested with Sca I and NgoM IV and the double right-border binary vector pMNDRBBin6 was digested with Hpa I and Xma I. pMNDRBBin6 carrying the gene Rxo1 was acquired by ligation of blunt-end and cohesive end. The results of PCR, restriction enzyme analysis and sequencing indicated that the Rxo1 gene had been cloned into pMNDRBBin6. This double right-border binary vector, named as pMNDRBBin6-Rxol, will play a role in breeding marker-free plants resistant to bacterial leaf streak of rice by genetic transformation.
基金Supported by National 863 Project of China (2002AA227011)Natural Science Foundation of Hubei Province (2003ABAI18)Natural Science Foundation of Shandong Province (ZR2010HQ054)~~
文摘[Objective] This study aimed to construct Brassica napus chloroplast multi- cistron double cross-over expression vector, to lay the foundation for the genetic engi- neering research of Brassica napus chloroplast. [Method] Two primers were designed based on the known Brassica napus chloroplast DNA sequences AF267640 and Z50868 in GenBank. By using PCR method, two Brassica napus L. chloroplast DNA fragments were obtained, which were named RbcL and ACCD. The two Brassica na- pus chloroplast DNA homologous fragments were then cloned into plasmid pMD18-T to obtain recombinant plasmid pHBM715. Tandem expression cassette harboring spectinomycin-resistant gene aadA, mannanase gene man and green fluorescent pro- tein gene gfp was cloned into the plasmid pHBM715, thereby constructing Brassica napus chloroplast multicistron double cross-over expression vector pHBM716, which was transformed into Escherichia coil for expression and identification. [Result] Plate qualitative analysis was conducted for the functional identification of expression cas- sette in the constructed Brassica napus chloroplast multicistron double cross-over ex- pression vector, results showed that the three genes of the same multicistron were all expressed in E. coil [Conclusion] This study successfully constructed Brassica napus chloroplast multicistron double cross-over expression vector, which laid the foundation for the genetic engineering of Brassica napus chloroplast.
文摘We have developed a 'double T-DNA' binary vector system for generating selectable marker-free transgenic plants by Agrobacterium-mediated transformation. The 'double T-DNA' binary vector pDLBRBbarm which carried two independent T-DNAs, one containing a selectable marker neomycin phosphotransferase (nptII) gene and the other a bargene, was constructed. Transgenic tobacco (Nicotiana tabacum L.) plants were then produced by Agrobacterium-mediated transformation with this vector. Frequency of the primary transformants co-integrated with npt II gene and bar gene was 59.2%. Segregation of two T-DNA regions was found in 3 out of 4 T-1 lines from co-transformed T-0 plants with nptII and bar PPT-resistant and kanamycin-sensitive plants were in approximate 19.5% of the T-1 plants. The result indicated that this 'double T-DNA' vector system could be a workable approach to generate transgenic plants free from selectable marker genes. Co-transformation of nptII gene and bar gene to plants with mixtures of Agrobacterium tumefaciens strains containing single T-DNA vectors was also tested. Frequency of co-transformed plants was 20.0%-47.7% and relatively low as compared with that of 'double T-DNA' vector system.
文摘In order to overcome the interference of the internal promoter in retroviral vector with a foreign DNA expression, we used the double-copy vector(DC vector) system to improve the effect of transduced gene expression.Human G-CSF cDNA was inserted into the Bgl Ⅱ site of the polyclonal sites within the U3 region of the 3'long terminal repeat(3'-LTR) in the vector(N2A). After being identified, the gene was transduced into Ψ-2 packaging cell line by using the electroporation method. Consequently, the gene was duplicated in the infected cells, and transferred to the 5'-LRT, and then placed outside the retroviral transcriptional unit. After two weeks the neomycin resistance positive colonies were grown in the G-418 medium. The supernatant virus was titred and then transferred to NIH3T3 cells. The G-CSF value of positive clone N2AG7-4 was up to 53. 3 U/106 cells. Southern blot and Northern blot analysis showed that the chimeric gene faithfully duplicated in the cells infected with the corresponding virus and generated two copies with one in each LTR.
基金the National Natural Science Foundation of China(Nos.61071152 and 61271316)the National Basic Research Program (973) of China (Nos.2010CB731403 and 2010CB731406)the National "Twelfth Five-Year" Plan for Science and Technology Support(No.2012BAH38 B04)
文摘Copy-paste forgery is a very common type of forgery in JPEG images.The tampered patch has always suffered from JPEG compression twice with inconsistent block segmentation.This phenomenon in JPEG image forgeries is called the shifted double JPEG(SDJPEG) compression.Detection of SDJPEG compressed image patches can make crucial contribution to detect and locate the tampered region.However,the existing SDJPEG compression tampering detection methods cannot achieve satisfactory results especially when the tampered region is small.In this paper,an effective SDJPEG compression tampering detection method utilizing both intra-block and inter-block correlations is proposed.Statistical artifacts are left by the SDJPEG compression among the magnitudes of JPEG quantized discrete cosine transform(DCT) coefficients.Firstly,difference 2D arrays,which describe the differences between the magnitudes of neighboring JPEG quantized DCT coefficients on the intrablock and inter-block,are used to enhance the SDJPEG compression artifacts.Then,the thresholding technique is used to deal with these difference 2D arrays for reducing computational cost.After that,co-occurrence matrix is used to model these difference 2D arrays so as to take advantage of second-order statistics.All elements of these co-occurrence matrices are served as features for SDJPEG compression tampering detection.Finally,support vector machine(SVM) classifier is employed to distinguish the SDJPEG compressed image patches from the single JPEG compressed image patches using the developed feature set.Experimental results demonstrate the efficiency of the proposed method.
文摘A technique based on the double Fourier series is developed to estimate the winds at different isobaric levels forthe limited area domain, 35°E to 140°E and 30°S to 40°N, using the observed winds at 850 hPa lcvcl for the month ofJune. For this purpose the wind field at a level under consideration is taken in the ratio form with that of 850 hPa level and the coefficients of the double Fouricr series are computed. These coefficients are subsequently used to computethe winds which are compared with the actual winds. The results of the double Fourier series technique are comparedwith those of the polynomial surface fitting method developed by Bavadekar and Khaladkar (1 992). The technique isalso applied for the daily wind data of 11. June, 1979 and the validation of the technique is tested for a few radiosondestations of india. The computed winds for these radiosonde stations arc quite close to observed winds.
文摘DFSM （doubly fed synchronous machine） presents several advantages such as efficiency improvement, weight reduction and increase of the utilization factor （kW/kg）. In this paper the authors focus on impact of the DFSM on the efficiency and machine weight in comparison to conventional synchronous generator with wound rotor. Different topologies of DFSM are briefly described and the different methods and models for performances prediction are presented.
文摘A modeling method of the support vector machine combined with matrix optics is considered; a complete new measurement model for double-four quadrant photoelectric detector is built. According to the analysis of the received light spot size and its motion with the changes of the defocusing amount of detector photosensitive surface and the detector position attitude in the optical path, a mathematic expression of photoelectrical conversion is given, which can be applicable to random setting position of the detector at any time. Based on least square support vector machine (LS SVM), the mapping relationship among the output signal linear characteristic parameters (zero neighborhood gradient and intercept), the defocusing amount of the detector and the installation position attitude angle is established. Thus, the multiple dimensional high accuracy measuring and adjusting control system can be left out, and adaptive measurement of the detector parameters can be realized. Compared with existed measurement model and method, the presented model has the advantages of more clear physical meaning, closer to work mechanism of detector, acquiring more complete sample data and wiping out the dead spots or bad spots in measurement. And the accuracy of displacement measurement is increased to 3?μm. At the same time, this measurement mode provides a technical shortcut for three-dimensional small angle measurement.
文摘Expression vector pE14 with double resistance to virus and insect was constructed by inserting CW-cp gene and Bt-toxin gene one by into T-DNA of the same binary vector pE3. Tobacco was then transformed with Agrbacterium tumefaciens (At)GV311-SE carrying PE14. Nopaline assay and PCR amplification confirmed that both CW-cp gene and Bt-toxin gene had been introduced into tobacco genome by T-DNA of PE3. Test attack with virus and demonstrated, in some of the transgenic plants, double resistance to both infection by CMV and damage by Manduca sexta.
文摘为了解决多功能视频编码(versatile video coding,VVC)标准下具有相同编码参数的视频双压缩检测方法准确率不高的问题,提出了一种基于编码单元(coding unit,CU)尺寸、划分模式和预测模式的检测方法。对待检测的视频进行多次编解码,分析并确定VVC流中与压缩编码次数密切相关的基础码流特征;以CU尺寸、划分模式和预测模式构建高级码流特征输入支持向量机完成视频的双压缩检测。实验结果表明,与对比文献的方法相比,所提方法的视频双压缩检测准确率有较大提升,平均准确率达到了95.82%。
文摘针对永磁同步电机下桥臂电流采样拓扑的无位置传感器控制系统,在低载波运行时受数字延时影响易导致位置估计精度低、电流纹波增大、稳定性差等问题,提出一种半周期计算、单采样双更新的空间矢量脉宽调制方法,有效减少系统数字延时。该方法在每个脉冲宽度调制(Pulse-Width Modulation,PWM)的载波过零处采样一次电流,并在采样结束后的半周期内完成目标电压矢量计算。考虑数字延时中电机转过的角度,将目标电压矢量在下一个PWM周期的前、后半周期输出,并分别补偿0.75 T s和1.25 T s等效延时(T s为PWM周期时间),以减小目标电压矢量与实际输出电压矢量之间的误差。将新方法应用于采用模型参考自适应速度观测器的永磁同步电机无位置传感器控制系统中,实验结果表明该方法位置估计精度高、电流纹波小,且可以稳定运行于额定负载、载波比为4.0的工况。
