| 为了保证全球列国转基因产物标识治理轨制的顺遂实行，针对转基因水稻检测办法中存在内尺度基因检测办法不同一、转基因棉花品系特异性检测办法有待完美等成绩，本文展开了相干针对性研究。1。合适转基因水稻定性定量PCR检测的内尺度基因的国际协同验证依据我们曾经树立的水稻内尺度基因Sucrose Phosphate Synthase (SPS)定性定量PCR检测系统，优化定性定量PCR引物、探针浓度、序列和荧光标志位点，法语论文题目，法语论文题目，树立了稳固高效的SPS定性和定量PCR检测体系，并组织展开了在国际间试验室内尺度基因检测办法的验证，该国际协同试验约请国际外7个国度12个试验室分离对水稻内尺度基因SPS停止验证。成果注解，SPS基因相符转基因产物检测须要的内尺度基因的三个特点，即种间特异性，种内非特异性和恒定的低拷贝数。协同试验验证SPS定性PCR检测办法的检测上限可以或许达到0。1%。定量PCR检测办法的检测上限和定量上限至多为23个拷贝，而且具有优越的反响效力和线性相干系数。采取SPS定量PCR办法对8个水稻盲样剖析的误差在5。22%到26。52%之间。是以解释SPS合适作为水稻的内尺度基因，合适用于对转基因水稻样品停止定性和定量PCR检测。2。依据我国曾经同意贸易化的转基因棉花Mon15985的5’端旁侧序列和正在同意贸易化中的转基因棉花Mon88913的3’端旁侧序列，设计引物和探针，经由过程优化定性定量PCR检测办法的引物探针浓度，PCR检测系统，树立了转基因棉花Mon15985和Mon88913的品系特异性定性和定量PCR检测办法，成果证实了这两种转基因棉花品系特异性定性PCR检测办法具有高特异性，敏锐度到达0。05%，定量PCR检测办法的检测上限和定量上限分离约10和17个拷贝。在此基本上，我们在试验室外部组织了5人的协同试验验证，成果注解这两种转基因棉花的品系特异性定量PCR检测办法具有高PCR扩增效力，高线性相干系数的特色。而且对5个浓度梯度的转基因棉花混杂样品的定量检测成果与其现实成果的误差在25%规模内。成果注解所树立的两种转基因棉花品系特异性PCR检测办法完整实用于这两种转基因棉花及其衍生物的判定和定量剖析。别的，我们依据已报导的两种转基因棉花Mon1445和Mon531的品系特异性检测办法和本研究树立的Mon15985和Mon88913品系特异性检测办法，树立四种转基因棉花(Mon88913，Mon15985，Mon1445和Mon531)复合品系特异性定性PCR检测办法。该复合定性PCR检测办法在转基因棉花检测中具有高特异性和低的检测上限(0。1%)等长处，进步了转基因棉花定性检测办法的效力。这些对于转基因产物标识治理轨制的顺遂实行供给了技巧支持。 Abstract: In order to ensure global nations transgene product identification management rail system performed smoothly. In view of the existing measures for the detection of transgenic rice in inner scale gene detection method different, transgenic cotton lines specific detection method to be perfect scores. In this thesis, the coherent for seminars. 1. Suitable transgenic rice qualitative and quantitative PCR detection of the inner scale gene International Co verification basis we have set up the rice in scale gene sucrose phosphate synthase (SPS) qualitative and quantitative PCR detection system, optimize the qualitative quantitative PCR primers, concentration of probe sequences and fluorescence marker loci, and establish a stable and efficient SPS qualitative and quantitative PCR detection system, organized and launched a laboratory scale gene detection method be verified in the international, the international collaborative test invited international seven nations 12 laboratory separation of rice in the scale of SPS gene stop verification. Results note that SPS gene conform to the three characteristics of the transgenic product tests require the inner scale gene, i.e. interspecific specificity, non specificity and constant low copy number. The detection limit of collaborative testing verification of SPS qualitative PCR detection method can reach 0. 1%. The detection limit and the upper limit of quantification of quantitative PCR detection method of up to 23 copies, but also has superior response effect and linear correlation coefficient. Take the SPS quantitative PCR method for the error analysis of the 8 rice samples in 5. 22% to 26. 52%. Is suitable to explain SPS as inner scale rice, suitable for transgenic rice sample for qualitative and quantitative detection of PCR. 2. Basis in China had agreed to trade of transgenic cotton Mon15985 5 'end flanking sequences and are agreed to trade in transgenic cotton Mon88913 3' end flanking sequence, primers and probes were designed, optimized through the process of qualitative and quantitative PCR detection method of primer probe concentration, PCR detection system, establish the transgenic cotton Mon15985 and Mon88913 strain specific of qualitative and quantitative PCR detection methods, the results confirm the the two transgenic cotton lines specific qualitative PCR detection method has high specificity and acuity reaches 0. 05%, the detection limit and the upper limit of quantification of quantitative PCR detection method of separation of about 10 and 17 copies. On this basis, we on the outside of the laboratory organized five collaborative experimental verification, the results note the two transgenic cotton lines specific quantitative PCR detection method with PCR amplification effect, high linear correlation coefficient characteristics. And the results of quantitative detection of transgenic cotton on the concentration gradient of the 5 hybrid samples and real results in 25% scale error. Two kinds of transgenic cotton lines with specific PCR detection methods set up a complete and practical annotation results in the two kinds of transgenic cotton and its derivatives determination and quantitative analysis. Else, we reported two transgenic cotton was systematically optimized and developed the strain specific detection method and the research basis to establish the Mon15985 and Mon88913 strains specificity detection measures, establish four kinds of Transgenic Cotton (Mon88913, Mon15985 was systematically optimized and developed composite strain specific qualitative PCR detection method. The composite qualitative PCR detection method with high specificity and low detection limit in the detection of Transgenic Cotton (0. 1%) other strengths, improve the efficiency of transgenic cotton qualitative detection method. The product identification management system on genetically modified and well implemented and provides the technical support. 目录: |
