Abstract:Abstract: To make up for the lack of traditional reference materials, a plasmid reference molecule was constructed for screening the components of genetically modified crops. First, the 454 bp RBCL gene, 236 bp CaMV35S promoter and 200 bp NOS terminator were amplified by PCR. Using overlap extension PCR technology the three amplified fragments were ligated to be a 849 bp recombinant. After cloning of the recombinant into pMD18-T vector, the positive recombinant plasmid pMD-RBCL-CaMV35S-NOS was successfully confirmed by PCR amplification and sequence analysis. Further substitution studies showed that the analysis results of the recombinant plasmid DNA were identical with those of the genomic DNA of the standard substance. Therefore, the recombinant plasmid standard molecule can be used as a positive reference material for preliminary screening of the components of genetically modified crops.
引用本文:
肖维威, 张 宝, 赵 卫, 朱 利, 吴清华. 用于筛查转基因作物成分的质粒标准分子的研制[J]. 生命科学研究, 2019, 23(3): 192-199.
XIAO Wei-wei, ZHANG Bao, ZHAO Wei, ZHU Li, WU Qing-hua. Developing a Standard Reference Molecule for Screening Genetically Modified Crop Components. Life Science Research, 2019, 23(3): 192-199.