利用引物探针序列构建转基因马铃薯检测阳性重组质粒及其验证  

Construction of Transgenic Potato Detection Positive Recombinant Plasmid by Primer Probe Sequence and Its Validation

作　　者：杨杰 李兰 戴莉 张江国 莫善明 徐新龙 YANG Jie;LI Lan;DAI Li(Technical Center of Alashankou Customs,Alashankou,Xinjiang 833418)

机构地区：[1]阿拉山口海关技术中心

出　　处：《安徽农业科学》2019年第23期213-218,共6页Journal of Anhui Agricultural Sciences

基　　金：新疆维吾尔自治区国际合作项目(20166012)

摘　　要：[目的]通过将特定转基因检测方法引物和探针序列重组拼接,并将重组序列连接至质粒中,验证其作为转基因检测阳性对照的可行性。[方法]通过搭桥PCR和全基因合成的方法制备阳性重组序列,验证阳性重组序列作为阳性对照品的可行性与阳性重组质粒的均一性、稳定性。[结果]通过将引物探针序列顺序拼接构建得到的阳性重组质粒的性状均一,在4和-20℃条件下均能稳定产生阳性结果。[结论]该种阳性重组质粒的构建方法能够满足转基因成分检测方法中阳性对照品的要求。[Objective]The primers and probe sequences of specific GMO detection method were recombined and spliced,and the recombinant sequences were linked to the plasmid.To verify its feasibility as a positive control for GMO detection.[Method]Positive recombinant sequences were prepared by overlap PCR and whole gene synthesis to verify the feasibility of using positive recombinant sequences as positive reference materials and the homogeneity and stability of positive recombinant plasmid.[Result]The positive recombinant plasmids constructed by sequential splicing of primer probe sequences were uniform in character and could produce stable positive results at 4℃and-20℃.[Conclusion]The construction method of the positive recombinant plasmid can meet the requirements of the positive control in the detection of GMO.

关 键 词：引物 探针序列 转基因检测 搭桥PCR 阳性重组质粒 

分 类 号：Q943.2[生物学—植物学]