It is first reported that plant young proembryos expressed exogenous reporter genes by electroporation. Young proembryos with 8-32 cells and globular proembryos with 250-400 cells could be isolated by enzymatic macera...It is first reported that plant young proembryos expressed exogenous reporter genes by electroporation. Young proembryos with 8-32 cells and globular proembryos with 250-400 cells could be isolated by enzymatic maceration combined with microdissection. After electroporation with GUS or GFP genes, the proembryos were cultured for 1 -2 d in KM8p medium. At the field strength of electroporation 500-1 500 V/cm, blue reaction of GUS or green fluorescence of GFP could be observed in the proembryos. The highest transient expression frequency of young proembryos (2.2%) was obtained at the field strength of 750 V/cm, whereas the highest frequency of globular proembryos (5.9%) was obtained at the field strength of 1 250 V/cm. Taking the proportion of transformed cells in the whole cells of proembryos as efficient transformation frequency, the efficient transformation frequency of the young proembryos was 7 times that of the globular proembryos.展开更多
Two cDNA libraries were constructed from microdissected 214 rice proembryos (2-3 d after pollination) and 121 just differentiating young embryos (3-5 d after pollination) respectively through RT_PCR technique. The pri...Two cDNA libraries were constructed from microdissected 214 rice proembryos (2-3 d after pollination) and 121 just differentiating young embryos (3-5 d after pollination) respectively through RT_PCR technique. The primary libraries had a total of 3.7×10 6 phages for the proembryos and a total of 2.5×10 6 phages for the just differentiating young embryos, in which 96% of the phages were recombinants. Insert sizes ranging from 400 bp to 3?500 bp were obtained. All of the above mentioned accorded with the general requirements of cDNA library construction.展开更多
文摘It is first reported that plant young proembryos expressed exogenous reporter genes by electroporation. Young proembryos with 8-32 cells and globular proembryos with 250-400 cells could be isolated by enzymatic maceration combined with microdissection. After electroporation with GUS or GFP genes, the proembryos were cultured for 1 -2 d in KM8p medium. At the field strength of electroporation 500-1 500 V/cm, blue reaction of GUS or green fluorescence of GFP could be observed in the proembryos. The highest transient expression frequency of young proembryos (2.2%) was obtained at the field strength of 750 V/cm, whereas the highest frequency of globular proembryos (5.9%) was obtained at the field strength of 1 250 V/cm. Taking the proportion of transformed cells in the whole cells of proembryos as efficient transformation frequency, the efficient transformation frequency of the young proembryos was 7 times that of the globular proembryos.
文摘Two cDNA libraries were constructed from microdissected 214 rice proembryos (2-3 d after pollination) and 121 just differentiating young embryos (3-5 d after pollination) respectively through RT_PCR technique. The primary libraries had a total of 3.7×10 6 phages for the proembryos and a total of 2.5×10 6 phages for the just differentiating young embryos, in which 96% of the phages were recombinants. Insert sizes ranging from 400 bp to 3?500 bp were obtained. All of the above mentioned accorded with the general requirements of cDNA library construction.
