上尚科技发布一系列针对远程无线传输的产品GPRS Serial Servers(SG6001智能型单串口GPRS／GSM无线串行服务器；SG6103内建八组数字I／O三端口GPRS／GSM无线串行服务器)，可透过GPRS／GSM无线通讯网路，对远程设备进行监控与管理。可应...上尚科技发布一系列针对远程无线传输的产品GPRS Serial Servers(SG6001智能型单串口GPRS／GSM无线串行服务器；SG6103内建八组数字I／O三端口GPRS／GSM无线串行服务器)，可透过GPRS／GSM无线通讯网路，对远程设备进行监控与管理。可应用于包括水位、温湿度及各种环境监控、煤气及石油流量测量、远程终端机、交通号志监控及车队管理等方面。展开更多
CRISPR-Cas9 system is now widely used to edit a target genome in animals and plants. Cas9 protein derived from Streptococcus pyogenes（Sp Cas9） cleaves double-stranded DNA targeted by a chimeric single-guide RNA（sg ...CRISPR-Cas9 system is now widely used to edit a target genome in animals and plants. Cas9 protein derived from Streptococcus pyogenes（Sp Cas9） cleaves double-stranded DNA targeted by a chimeric single-guide RNA（sg RNA）. For plant genome editing, Agrobacterium-mediated T-DNA transformation has been broadly used to express Cas9 proteins and sg RNAs under the control of Ca MV 35 S and U6/U3 promoter, respectively. We here developed a simple and high-throughput binary vector system to clone a 19 20 bp of sg RNA, which binds to the reverse complement of a target locus, in a large T-DNA binary vector containing an Sp Cas9 expressing cassette. Twostep cloning procedures：（1） annealing two target-specific oligonucleotides with overhangs specific to the Aar I restriction enzyme site of the binary vector; and（2） ligating the annealed oligonucleotides into the two Aar I sites of the vector, facilitate the high-throughput production of the positive clones. In addition, Cas9-coding sequence and U6/U3 promoter can be easily exchanged via the GatewayTMsystem and unique Eco RI/Xho I sites on the vector, respectively. We examined the mutation ratio and patterns when we transformed these constructs into Arabidopsis thaliana and a wild tobacco, Nicotiana attenuata. Our vector system will be useful to generate targeted large-scale knock-out lines of model as well as non-model plant.展开更多
文摘上尚科技发布一系列针对远程无线传输的产品GPRS Serial Servers(SG6001智能型单串口GPRS／GSM无线串行服务器；SG6103内建八组数字I／O三端口GPRS／GSM无线串行服务器)，可透过GPRS／GSM无线通讯网路，对远程设备进行监控与管理。可应用于包括水位、温湿度及各种环境监控、煤气及石油流量测量、远程终端机、交通号志监控及车队管理等方面。
基金supported by Institute for Basic Science (IBS-R021-D1)
文摘CRISPR-Cas9 system is now widely used to edit a target genome in animals and plants. Cas9 protein derived from Streptococcus pyogenes（Sp Cas9） cleaves double-stranded DNA targeted by a chimeric single-guide RNA（sg RNA）. For plant genome editing, Agrobacterium-mediated T-DNA transformation has been broadly used to express Cas9 proteins and sg RNAs under the control of Ca MV 35 S and U6/U3 promoter, respectively. We here developed a simple and high-throughput binary vector system to clone a 19 20 bp of sg RNA, which binds to the reverse complement of a target locus, in a large T-DNA binary vector containing an Sp Cas9 expressing cassette. Twostep cloning procedures：（1） annealing two target-specific oligonucleotides with overhangs specific to the Aar I restriction enzyme site of the binary vector; and（2） ligating the annealed oligonucleotides into the two Aar I sites of the vector, facilitate the high-throughput production of the positive clones. In addition, Cas9-coding sequence and U6/U3 promoter can be easily exchanged via the GatewayTMsystem and unique Eco RI/Xho I sites on the vector, respectively. We examined the mutation ratio and patterns when we transformed these constructs into Arabidopsis thaliana and a wild tobacco, Nicotiana attenuata. Our vector system will be useful to generate targeted large-scale knock-out lines of model as well as non-model plant.
