[Objective] This study aimed to investigate the major contributors to the measurement uncertainty in quantitative analysis of genetically modified ingredients and improve the quality of quantitative detection of genet...[Objective] This study aimed to investigate the major contributors to the measurement uncertainty in quantitative analysis of genetically modified ingredients and improve the quality of quantitative detection of genetically modified components. [Method] The content of Ca MV35 S promoter( parameter) in GTS40-3-2 soybean powder samples was measured to estimate the measurement uncertainty preliminarily. [Result]Type A uncertainty( uA),type B uncertainty(uB) and combined standard uncertainty( uC) were 0. 0 004,0. 002 and 0. 002,respectively. At a confidence level of p = 95% and freedom degree of Veff= 3 251,coverage factor k = 1. 96,expanded uncertainty U = 0. 004. The final measurement result was C = 0. 028 ± 0. 004,which was close to the conventional true value(0. 03).Thus,the measurement uncertainty was relatively small,indicating a high quality of measurement. In this study,uncertainty evaluation indicated that the deviation of micro liquid transfer made the greatest contribution to the measurement uncertainty. [Cluclusion]The deviation of micro liquid transfer should be reduced to improve the quality of measurement.展开更多
The gene Pi-d2,conferring gene-for-gene resistance to the Chinese blast strain ZB15,was isolated from a rice variety(Digu) by the map-based cloning strategy.Here,we constructed a control plasmid pZH01-pi-d2tp309(pZH01...The gene Pi-d2,conferring gene-for-gene resistance to the Chinese blast strain ZB15,was isolated from a rice variety(Digu) by the map-based cloning strategy.Here,we constructed a control plasmid pZH01-pi-d2tp309(pZH01-tp309) and three different expression constructs,pCB-Pi-d25.3kb(pCB5.3kb),pCB-Pi-d26.3kb(pCB6.3kb) and pZH01-Pi-d22.72kb(pZH01-2.72kb) of Pi-d2,driven by Pi-d2 gene's own promoter or CaMV35S promoter.These constructs were separately introduced into japonica rice varieties Lijiangxintuanhegu,Taipei 309,Nipponbare and Zhonghua 9 through Agrobacterium-mediated transformation.A total of 150 transgenic rice plants were obtained from the regenerated calli selected on hygromycin.PCR,RT-PCR and Southern-blotting assay showed that the gene of interest had been integrated into rice genome and stably inherited.Thirty-five transgenic lines independently derived from T1 progeny were inoculated with the rice blast strain ZB15.Transformants exhibited resistance to rice blast at various levels.The lesions on the transgenic plant leaves were less severe than those on the controls and the resistance level of transgenic plants harboring the gene of interest from three vectors had no difference.The own promoter of Pi-d2,about 2.2 kb or 3.2 kb,had the similar promoter function as CaMV35S.Field evaluation for three successive years supported the results of artificial trial,and some lines with high resistance to rice leaf blast and neck blast were obtained.展开更多
Xylem-specific promoter could regulate efficient expression of foreign genes in xylem. Deletion analysis method was applied to obtain 5' deletion promoter fragments MU2 andM U4 with different distances from transc...Xylem-specific promoter could regulate efficient expression of foreign genes in xylem. Deletion analysis method was applied to obtain 5' deletion promoter fragments MU2 andM U4 with different distances from transcription start site of xylem-specific promoter MDCesA P through PCR procedure. Then CaM V35 S promoter in plant expression vector pB I121 was replaced by amplified fragments. The recombinant plasmids fused with corresponding gene segments and GUS reporter gene were obtained and transformed into Agrobacterium. The results of the tobacco transient transformation test showed that MU2 andM U4 had the promoter function,and both of their activity were significantly higher than that of CaM V35 S promoter,and they had the characteristics of xylem tissue specificity as well. Research on xylem-specific promoter structure and function could lay a foundation for the determination of necessary components to the xylem tissue specificity and cis-acting elements with induction activity,so as to enable the regulation of efficient expression of exogenous genes in specific time and specific tissues in plants through these cis-acting elements.展开更多
基金Supported by Project of Standardized Technology System of Sichuan Bureau of Quality and Technical Supervision(ZYBZ2013-39)
文摘[Objective] This study aimed to investigate the major contributors to the measurement uncertainty in quantitative analysis of genetically modified ingredients and improve the quality of quantitative detection of genetically modified components. [Method] The content of Ca MV35 S promoter( parameter) in GTS40-3-2 soybean powder samples was measured to estimate the measurement uncertainty preliminarily. [Result]Type A uncertainty( uA),type B uncertainty(uB) and combined standard uncertainty( uC) were 0. 0 004,0. 002 and 0. 002,respectively. At a confidence level of p = 95% and freedom degree of Veff= 3 251,coverage factor k = 1. 96,expanded uncertainty U = 0. 004. The final measurement result was C = 0. 028 ± 0. 004,which was close to the conventional true value(0. 03).Thus,the measurement uncertainty was relatively small,indicating a high quality of measurement. In this study,uncertainty evaluation indicated that the deviation of micro liquid transfer made the greatest contribution to the measurement uncertainty. [Cluclusion]The deviation of micro liquid transfer should be reduced to improve the quality of measurement.
基金supported by the Excellent Doctor Paper Foundation of the Ministry of Education of China (Grant No.200054)the Program for Innovative Research Team in University of China (Grant No.NCET-04-0907)the Program for New Century Excellent Talent in University of China (Grant No.IRT0453)
文摘The gene Pi-d2,conferring gene-for-gene resistance to the Chinese blast strain ZB15,was isolated from a rice variety(Digu) by the map-based cloning strategy.Here,we constructed a control plasmid pZH01-pi-d2tp309(pZH01-tp309) and three different expression constructs,pCB-Pi-d25.3kb(pCB5.3kb),pCB-Pi-d26.3kb(pCB6.3kb) and pZH01-Pi-d22.72kb(pZH01-2.72kb) of Pi-d2,driven by Pi-d2 gene's own promoter or CaMV35S promoter.These constructs were separately introduced into japonica rice varieties Lijiangxintuanhegu,Taipei 309,Nipponbare and Zhonghua 9 through Agrobacterium-mediated transformation.A total of 150 transgenic rice plants were obtained from the regenerated calli selected on hygromycin.PCR,RT-PCR and Southern-blotting assay showed that the gene of interest had been integrated into rice genome and stably inherited.Thirty-five transgenic lines independently derived from T1 progeny were inoculated with the rice blast strain ZB15.Transformants exhibited resistance to rice blast at various levels.The lesions on the transgenic plant leaves were less severe than those on the controls and the resistance level of transgenic plants harboring the gene of interest from three vectors had no difference.The own promoter of Pi-d2,about 2.2 kb or 3.2 kb,had the similar promoter function as CaMV35S.Field evaluation for three successive years supported the results of artificial trial,and some lines with high resistance to rice leaf blast and neck blast were obtained.
基金Supported by National Science Foundation of China(31000305)Fund from Education Department of Hebei Province(QN2015182)+1 种基金Innovation Fund for Forestry Discipline of Hebei Agricultural University(LXXK2014-1)Fund for Overseas Research and Study by Young and Middle-aged Backbone Teachers in Hebei Agriculture University(JWYX2015)
文摘Xylem-specific promoter could regulate efficient expression of foreign genes in xylem. Deletion analysis method was applied to obtain 5' deletion promoter fragments MU2 andM U4 with different distances from transcription start site of xylem-specific promoter MDCesA P through PCR procedure. Then CaM V35 S promoter in plant expression vector pB I121 was replaced by amplified fragments. The recombinant plasmids fused with corresponding gene segments and GUS reporter gene were obtained and transformed into Agrobacterium. The results of the tobacco transient transformation test showed that MU2 andM U4 had the promoter function,and both of their activity were significantly higher than that of CaM V35 S promoter,and they had the characteristics of xylem tissue specificity as well. Research on xylem-specific promoter structure and function could lay a foundation for the determination of necessary components to the xylem tissue specificity and cis-acting elements with induction activity,so as to enable the regulation of efficient expression of exogenous genes in specific time and specific tissues in plants through these cis-acting elements.
