定性PCR结合核酸斑点杂交技术检测转基因番茄研究

In recent years the development of transgenic technology has cause great concern in society, especially the safety and rationality of this technology. Thus, establishment of the genetically modified crops detection method is particularly urgent. Tomato transgenic technology also has get great progress recently. China is an important tomato import and export country, it is necessary to explore an efficient tomato transgenic detection system. In the study, the traditional qualitative PCR and nucleic acid spots hybrid technology have been used to detect the transgenic components in samples from tomato Huafan No.1 and HeZuo903, using plasmid PBI121 as the positive control. The exogenous genes 35s promoter, NOS terminator, report gene NPTII, cross-linked structure of 35s and tomato gene EFE (35 s-EFE) (which are commonly used in transgenic plants); and the tomato reference gene Lat52 were studied. For each gene, 2-3 sequence specific primer pairs were designed, and the optimal was selected according to the actual PCR. All of the four exogenous genes and reference gene could be successful amplified in the samples from Huafan No.1, while only the reference gene Lat52 could be detected in HeZuo903. The repeatability of the detection is good, and sensitivity is high. The results of these two methods complement with each other. Multiple genes could be detected by nucleic acid spots hybrid at the same time, which increases the detection efficiency. The results of this study provide a reference for tomato transgenic detection.