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Novel reference gene, high-mobility-group protein I/Y, used in qualitative and real-time quantitative polymerase chain reaction detection of transgenic rapeseed cultivars

文献类型： 外文期刊

作者： Weng, HB 1 ; Yang, LT 2 ; Liu, ZL 2 ; Ding, JY 3 ; Pan, AH; Zhang, DB;

作者机构： 1.Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China

2.Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China; Shanghai Acad Agr Sci, Shanghai Key Lab Agr Genet & Breeding, Agro Biotech Res Ctr, Shanghai 201106, Peoples R China; Shanghai Teachers Univ, Life & Environm Sci Coll, Shanghai 200234, Peoples R China; Nanjing Univ, Dept Biol Sci & Technol, Nanjing 210093, Peoples R China

3.Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China; Shanghai Acad Agr Sci, Shanghai Key Lab Agr Genet & Breeding, Agro Biotech Res Ctr, Shanghai 201106, Peoples R China; Shanghai Teach

期刊名称：JOURNAL OF AOAC INTERNATIONAL （ 影响因子：1.913； 五年影响因子：1.754 ）

ISSN： 1060-3271

年卷期： 2005 年 88 卷 2 期

页码：

收录情况： SCI

摘要： With the development of transgenic crops, regulations to label the genetically modified organisms (GMOs) and their derived products have been issued in many countries. Polymerase chain reaction (PCR) methods are thought to be reliable and useful techniques for qualitative and quantitative detection of GMOs. These methods are generally needed to amplify the transgene and compare the amplified results with that of a corresponding reference gene to get the reliable results. Specific primers were developed for the rapeseed (Brassica napus), high-mobility-group protein I/Y(HMG-I/Y) single-copy gene and PCR cycling conditions suitable for the use of this sequence as an endogenous reference gene in both qualitative and quantitative PCR assays. Both methods were assayed with 15 different rapeseed varieties, and identical amplified products were obtained with all of them. No amplification was observed when templates were the DNA samples from the other species of Brassica genus or other species, such as broccoli, stem mustard, cauliflower, Chinese cabbage, cabbage, sprouts, Arabidopsis thaliana, carrot, tobacco, soybean, mung bean, tomato, pepper, eggplant, plum, wheat, maize, barley, rice, lupine, and sunflower. This system was specific for rapeseed. Limits of detection and quantitation in qualitative and quantitative PCR systems were about 13 pg DNA (about 10 haploid genomes) and about 1.3 pg DNA (about 1 haploid genome), respectively. To further test the feasibility of this HMG-I/Y gene as an endogenous reference gene, samples containing transgenic rapeseed GT73 with the inserted glyphosate oxidoreductase (GOX) gene were quantitated. These demonstrated that the endogenous PCR detection systems were applicable to the qualitative and quantitative detection of transgenic rapeseed.

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