[Objective] The aim was to construct a plasmid reference molecule (PRM) for detection of transgenic soybean MON89788. [Method] the lectin gene sequence,3'-junction and 5'-junction sequence between host plant D...[Objective] The aim was to construct a plasmid reference molecule (PRM) for detection of transgenic soybean MON89788. [Method] the lectin gene sequence,3'-junction and 5'-junction sequence between host plant DNA integrated DNA of MON89788 soybean were amplified independently,and the three fragments were cloned into the cloning vector pMD18-T in order through molecular manipulation method to construct pMD-LM3M5,the applicability of the constructed novel PRM was tested. [Result] Sequencing confirmation result showed that the PRM was 3 700 bp in length,containing 1 029 bp of recombined DNA fragment. The limits of qualitative detection of the PRM were 10 copies. [Conclusion] The PRM constructed in this study was suitable for the identification of MON89788 event.展开更多
Diapause hormone (DH) is a neurohormone which is secreted from suboesophageal ganglion and responsible for induction of embryonic diapause in Bombyx mori.Here,using bioassay,we present evidence that Has-SGNPⅠ,which c...Diapause hormone (DH) is a neurohormone which is secreted from suboesophageal ganglion and responsible for induction of embryonic diapause in Bombyx mori.Here,using bioassay,we present evidence that Has-SGNPⅠ,which clearly exhibited diapause egg inducing activity as does Bom-DH,is likely to be DH.We reported the molecular characterization of the Helicoverpa assulta diapause hormone analog by polymerase chain reaction.A 665?bp fragment containing the entire DH-like gene was isolated and characterized.DH-like gene comprised two exons interspersed by a single intron,and the consensus sequence for splicing junction is identified at the exon/intron boundary.The result shows that DH-like gene is localized in the genome of Helicoverpa assulta.展开更多
Objective: To directionally clone the ompl gene fromChlamydia trachomatis (Ct) F genotype onto a plasmid vectorfor constructing a rudimentary DNA vaccine. Methods: The complete ompl gene from genomic DNA of CtF genoty...Objective: To directionally clone the ompl gene fromChlamydia trachomatis (Ct) F genotype onto a plasmid vectorfor constructing a rudimentary DNA vaccine. Methods: The complete ompl gene from genomic DNA of CtF genotype wild species was amplified with primers designedby computer. The recombinant gene was obtained byrestriction enzyme cutting, linking the gene with the plasmidvector in vitro, transforming the recombinant gene intobacteria, and extracting the DNA from the bacteria. Results: DNA extracted from the bacteria was composed ofthe ompl gene and plasmid, which is identified by threemethods of singular restrictive enzyme cutting, doublerestrictive enzyme cutting and PCR. Conclusion: Cloning of the ompl gene from the Ct Fgenotype means that a rudimentary DNA vaccine wassuccessfully constructed.展开更多
基金Supported by Major Projects of Cultivating New Varieties by Trans-genic Technology (2008ZX08012-001)~~
文摘[Objective] The aim was to construct a plasmid reference molecule (PRM) for detection of transgenic soybean MON89788. [Method] the lectin gene sequence,3'-junction and 5'-junction sequence between host plant DNA integrated DNA of MON89788 soybean were amplified independently,and the three fragments were cloned into the cloning vector pMD18-T in order through molecular manipulation method to construct pMD-LM3M5,the applicability of the constructed novel PRM was tested. [Result] Sequencing confirmation result showed that the PRM was 3 700 bp in length,containing 1 029 bp of recombined DNA fragment. The limits of qualitative detection of the PRM were 10 copies. [Conclusion] The PRM constructed in this study was suitable for the identification of MON89788 event.
文摘Diapause hormone (DH) is a neurohormone which is secreted from suboesophageal ganglion and responsible for induction of embryonic diapause in Bombyx mori.Here,using bioassay,we present evidence that Has-SGNPⅠ,which clearly exhibited diapause egg inducing activity as does Bom-DH,is likely to be DH.We reported the molecular characterization of the Helicoverpa assulta diapause hormone analog by polymerase chain reaction.A 665?bp fragment containing the entire DH-like gene was isolated and characterized.DH-like gene comprised two exons interspersed by a single intron,and the consensus sequence for splicing junction is identified at the exon/intron boundary.The result shows that DH-like gene is localized in the genome of Helicoverpa assulta.
文摘Objective: To directionally clone the ompl gene fromChlamydia trachomatis (Ct) F genotype onto a plasmid vectorfor constructing a rudimentary DNA vaccine. Methods: The complete ompl gene from genomic DNA of CtF genotype wild species was amplified with primers designedby computer. The recombinant gene was obtained byrestriction enzyme cutting, linking the gene with the plasmidvector in vitro, transforming the recombinant gene intobacteria, and extracting the DNA from the bacteria. Results: DNA extracted from the bacteria was composed ofthe ompl gene and plasmid, which is identified by threemethods of singular restrictive enzyme cutting, doublerestrictive enzyme cutting and PCR. Conclusion: Cloning of the ompl gene from the Ct Fgenotype means that a rudimentary DNA vaccine wassuccessfully constructed.