masD和bamA基因分别是烷烃和芳烃厌氧降解的关键基因。本研究建立了这两种基因的SYBR Green Ⅰ实时荧光定量PCR检测方法。通过参考相关石油烃厌氧降解菌株的Gen Bank序列,利用Primer express软件设计烷烃和芳烃厌氧降解基因的扩增引物ma...masD和bamA基因分别是烷烃和芳烃厌氧降解的关键基因。本研究建立了这两种基因的SYBR Green Ⅰ实时荧光定量PCR检测方法。通过参考相关石油烃厌氧降解菌株的Gen Bank序列,利用Primer express软件设计烷烃和芳烃厌氧降解基因的扩增引物masD-f、masD-r和bamA-f、bamA-r。经过常规PCR扩增分别得到片段大小为389和354 bp扩增产物,经测序并在NCBI数据库查询,确定为masD和bamA片段。通过实时荧光定量PCR构建测定这两种基因的标准曲线。优化后的扩增体系(25μL)为:12.5μL 2×Trans Start Top Green qPCR Super Mix,引物浓度为0. 2μmol/L,masD和bamA基因最适退火温度分别为52℃和56℃。建立的两种基因的实时荧光定量PCR检测方法具有非常好的重复性,其灵敏度比传统PCR技术高100倍。对于氧化石墨烯促进石油烃的厌氧降解体系中厌氧基因的定量检测显示,添加不同浓度的石墨烯均促进了bamA拷贝数的增加,但对masD的拷贝数无显著影响。展开更多
masD和bamA是控制石油烃厌氧降解的关键基因,利用实时荧光定量PCR技术检测masD和bamA基因具有简便快速和易操作等优点.但目前所用方法存在扩增效率低,方法灵敏度较差的问题.本文根据引物设计原则,利用Allele ID6软件重新设计了扩增masD...masD和bamA是控制石油烃厌氧降解的关键基因,利用实时荧光定量PCR技术检测masD和bamA基因具有简便快速和易操作等优点.但目前所用方法存在扩增效率低,方法灵敏度较差的问题.本文根据引物设计原则,利用Allele ID6软件重新设计了扩增masD和bamA的实时荧光定量PCR引物,将质粒DNA进行8次10倍梯度稀释后构建实时荧光定量PCR标准曲线.优化后的体系(20μL)为:FastStart Essential DNA Green Master 10.0μL,上下游引物各0.4μL,RNase-Free Water 4.2μL,5.0μL DNA模板.利用新设计的引物扩增masD和bamA基因的最适退火温度分别为61℃和57℃.优化后的检测方法扩增效率提高至97.5%和71.2%,比文献报道的方法提高了7.6%—44.5%,具有更高的重复性和灵敏度.利用设计的引物对陕北5个地区石油污染土壤中的masD和bamA基因进行定量检测结果表明,石油污染土壤中普遍存在着控制石油烃厌氧降解的关键基因,所测定的土壤中bamA降解基因的拷贝数远高于masD降解基因.展开更多
The micro-aggregated size distribution (MASD) of surface soil is an important parameter for modelling dust emission. However, there is no dataset of MASDs of all sur-face soil types in north China. The MASDs are here ...The micro-aggregated size distribution (MASD) of surface soil is an important parameter for modelling dust emission. However, there is no dataset of MASDs of all sur-face soil types in north China. The MASDs are here pre-sented, measured by dry sieving, for typical surface soil sam-ples, including sandy soil, gravelly sand soil, gravelly loam soil, loam soil and silt loam soil, collected from sandy deserts, Gobi deserts, oases, farmlands in steppe regions and steppe areas in north China. The MASDs of various surface soil types exhibit a combination of several log-normal distribu-tions of five separated sizes with mean mass median diame-ters (MMDs) of 90, 210, 390, 600 and 980 mm, respectively, and mean standard deviations (SDs) of 1.25, 1.40, 1.25, 1.35 and 1.25 respectively. The log-normal distributions corre-spond to very fine sand, fine sand, medium sand, coarse sand and very coarse sand population. On the basis of characteri-zation of the retrieved MASDs of various surface soil types in north China, dust emission fluxes are modelled by a dust production model (DPM model). It is shown that dust emis-sion has been significantly influenced by MASDs. Fine sand and very fine sand are always associated with the highest dust emission fluxes. Emission fluxes of the medium sand, gravelly sand soil, gravelly loam soil and loam soil are lower than those of very fine sand and fine sand, but larger than those of the coarse sand. The differences in dust emission fluxes vary among the different soil types from 101 to 103 mg·m-2·s-1. Dust emission fluxes from sandy deserts and farmlands covered with sand sheets in north China rang from 101 to 104 mg·m-2·s-1 while those from Gobi deserts, farmlands and steppes with gravelly desertification range from 101 to 102 mg·m-2· s-1. The modelled results indicate that deserts and farmlands with sand are the major dust sources in north China.展开更多
文摘masD和bamA基因分别是烷烃和芳烃厌氧降解的关键基因。本研究建立了这两种基因的SYBR Green Ⅰ实时荧光定量PCR检测方法。通过参考相关石油烃厌氧降解菌株的Gen Bank序列,利用Primer express软件设计烷烃和芳烃厌氧降解基因的扩增引物masD-f、masD-r和bamA-f、bamA-r。经过常规PCR扩增分别得到片段大小为389和354 bp扩增产物,经测序并在NCBI数据库查询,确定为masD和bamA片段。通过实时荧光定量PCR构建测定这两种基因的标准曲线。优化后的扩增体系(25μL)为:12.5μL 2×Trans Start Top Green qPCR Super Mix,引物浓度为0. 2μmol/L,masD和bamA基因最适退火温度分别为52℃和56℃。建立的两种基因的实时荧光定量PCR检测方法具有非常好的重复性,其灵敏度比传统PCR技术高100倍。对于氧化石墨烯促进石油烃的厌氧降解体系中厌氧基因的定量检测显示,添加不同浓度的石墨烯均促进了bamA拷贝数的增加,但对masD的拷贝数无显著影响。
文摘masD和bamA是控制石油烃厌氧降解的关键基因,利用实时荧光定量PCR技术检测masD和bamA基因具有简便快速和易操作等优点.但目前所用方法存在扩增效率低,方法灵敏度较差的问题.本文根据引物设计原则,利用Allele ID6软件重新设计了扩增masD和bamA的实时荧光定量PCR引物,将质粒DNA进行8次10倍梯度稀释后构建实时荧光定量PCR标准曲线.优化后的体系(20μL)为:FastStart Essential DNA Green Master 10.0μL,上下游引物各0.4μL,RNase-Free Water 4.2μL,5.0μL DNA模板.利用新设计的引物扩增masD和bamA基因的最适退火温度分别为61℃和57℃.优化后的检测方法扩增效率提高至97.5%和71.2%,比文献报道的方法提高了7.6%—44.5%,具有更高的重复性和灵敏度.利用设计的引物对陕北5个地区石油污染土壤中的masD和bamA基因进行定量检测结果表明,石油污染土壤中普遍存在着控制石油烃厌氧降解的关键基因,所测定的土壤中bamA降解基因的拷贝数远高于masD降解基因.
文摘The micro-aggregated size distribution (MASD) of surface soil is an important parameter for modelling dust emission. However, there is no dataset of MASDs of all sur-face soil types in north China. The MASDs are here pre-sented, measured by dry sieving, for typical surface soil sam-ples, including sandy soil, gravelly sand soil, gravelly loam soil, loam soil and silt loam soil, collected from sandy deserts, Gobi deserts, oases, farmlands in steppe regions and steppe areas in north China. The MASDs of various surface soil types exhibit a combination of several log-normal distribu-tions of five separated sizes with mean mass median diame-ters (MMDs) of 90, 210, 390, 600 and 980 mm, respectively, and mean standard deviations (SDs) of 1.25, 1.40, 1.25, 1.35 and 1.25 respectively. The log-normal distributions corre-spond to very fine sand, fine sand, medium sand, coarse sand and very coarse sand population. On the basis of characteri-zation of the retrieved MASDs of various surface soil types in north China, dust emission fluxes are modelled by a dust production model (DPM model). It is shown that dust emis-sion has been significantly influenced by MASDs. Fine sand and very fine sand are always associated with the highest dust emission fluxes. Emission fluxes of the medium sand, gravelly sand soil, gravelly loam soil and loam soil are lower than those of very fine sand and fine sand, but larger than those of the coarse sand. The differences in dust emission fluxes vary among the different soil types from 101 to 103 mg·m-2·s-1. Dust emission fluxes from sandy deserts and farmlands covered with sand sheets in north China rang from 101 to 104 mg·m-2·s-1 while those from Gobi deserts, farmlands and steppes with gravelly desertification range from 101 to 102 mg·m-2· s-1. The modelled results indicate that deserts and farmlands with sand are the major dust sources in north China.
