中文摘要：本发明提供了一种多种转基因元件同时快速检测的方法，包括如下步骤：步骤1、多壁碳纳米管/氧化石墨烯纳米带MWCNTs/GONRs分散液的制备；步骤2、绿色碲化镉量子点溶液和红色碲化镉量子点溶液的制备；步骤3、碲化镉量子点?捕获探针分散液的制备；步骤4、荧光共振能量转移传感体系的构建；步骤5、标准曲线的构建；步骤6、基于荧光共振能量转移传感体系对转基因大豆的检测。该传感器的制备是基于DNA互补配对杂交特异性作用所构建的，因而相对于免疫反应而言有与目标分子结合力强、特异性高、长时间暴露不容易变性、对周围环境要求不高等特点。
外文摘要：NOVELTY - Detecting variety of transgenic components at same time involves preparing multi-walled carbon nanotubes (MWCNTs)/graphene nanotubes (GONRs) dispersions, preparing green cadmium telluride quantum dot solution and red cadmium telluride quantum dot solution, preparing cadmium telluride quantum dot-capture probe dispersion, where green cadmium telluride quantum dot solution and the red cadmium telluride quantum dot solution are ultrasonically dispersed, and the pH value of the solution of the green cadmium telluride quantum dot is adjusted by hydrochloric acid (HCl) to obtain mixed solution A.
USE - Method for detecting variety of transgenic components at same time (claimed).
ADVANTAGE - The method enables to detect variety of transgenic components at same time with strong ability to bind to target molecule and high specificity.
DETAILED DESCRIPTION - Detecting variety of transgenic components at same time involves preparing multi-walled carbon nanotubes (MWCNTs)/graphene nanotubes (GONRs) dispersions, preparing green cadmium telluride quantum dot solution and red cadmium telluride quantum dot solution, preparing cadmium telluride quantum dot-capture probe dispersion, where green cadmium telluride quantum dot solution and the red cadmium telluride quantum dot solution are ultrasonically dispersed, and the pH value of the solution of the green cadmium telluride quantum dot is adjusted by hydrochloric acid (HCl) to obtain mixed solution A. The pH of the red cadmium telluride quantum dot solution is adjusted by HCl to obtain the mixed solution B. The mixed solution A is added with 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride solution, followed by adding N-hydroxysuccinimide solution, shaking uniformly, adding the probe of the terminator, and reacting at room temperature to obtain the green cadmium telluride quantum dot-capture probe, denoted as NP-gQDs. A solution of 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride is added to the mixed solution B, and the mixture is uniformly shaken. N-hydroxysuccinimide solution is added and the mixture is added with promoter of the probe The reaction is carried out at room temperature to obtain a red cadmium telluride quantum dot-capture probe, denoted as PPrQDs followed bywashing with ethanol and centrifuging. Finally, NP-gQDs and PPrQDs are dispersed in Tris-HCl buffer to obtain NPgQDs dispersion and PP-rQDs dispersion respectively. The MWCNTs/GONRs dispersion is mixed with NP-gQDs solution and PP-rQDs solution and dispersed by ultrasonic to obtain the mixture and after standing, the fluorescence intensity is measured. The concentration of the terminal NOS solution and the promoter P35s solution are added to the mixed solution, and the fluorescence recovery degree is measured after ultrasonic and standing. The standard curve is constructed. The DNA solution of the transgenic soybean is extracted by the plant DNA extraction kit. The DNA of transgenic soybean is then added to the fluorescence resonance energy transfer sensing system constructed, and the fluorescence recovery intensity is compared with the standard curve to conclude whether NOS and P35s are present or not.
TF   TECHNOLOGY FOCUS - BIOTECHNOLOGY - Preferred Condition: NP-gQDs are prepared by using green cadmium telluride quantum dot solution, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride solution. The volume ratio of N-hydroxysuccinimide solution to capture probe stock is 42: 2: 2: 1. PP-rQDs are prepared by using red cadmium telluride quantum dot solution, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride solution. The volume ratio of N-hydroxysuccinimide solution to capture probe stock is 42: 2: 2: 1. The NP-gQDs and PPrQDs are prepared, where concentration of the 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride solution used is 400 mM, concentration of the N-hydroxysuccinimide solution used is 200 mM, concentration of the capture probe stock used is 100 mu M. The concentration of HCl used is 10 M, the adjusted pH is 5, the reaction time at room temperature is 40 min. The concentration of the Tris-HCl buffer solution used is 10 mM, pH is 7.4. The volume ratio of the Tris-HCl buffer solution used to the green cadmium telluride quantum dot solution used is 47:42. The volume ratio of the Tris-HCl buffer solution used to the red cadmium telluride quantum dot solution used is 47:42. The MWCNTs / GONRs dispersion used is 1.5 mg/mL, the volume ratio of MWCNTs/GONRs dispersion, NP-gQDs dispersion and PP-gQDs dispersion is 1: 750: 750. The concentration of the terminator NOS solution and the promoter P35s is 15: 15: 4 in the MWCNTs / GONRs dispersion, and the concentrations of the terminator NOS solution and the promoter P35s were 0 to 1000 nM. The ultrasonic time is 5 min and the standing time is 5 min. The DNA solution extracted from the transgenic soybean is 5: 1 in volume ratio to the MWCNTs / GONRs dispersion. Preferred Composition: The DNA sequences of promoter P35s, terminator NOS, promoter of the probe H2N and terminator of the probe H2N have 25, 25, 25 and 25 nucleobases (SEQ ID NOS: 1-4), which are ggcagaggcatcttcaacgatggcc, gattagagtcccgcaattatacatt, ggccatcgttgaagatgcctctgcc, aatgattaattgcgggactctaatc.
 
主权项：多种转基因元件同时快速检测的方法，其特征在于，包括如下步骤：步骤1、多壁碳纳米管/氧化石墨烯纳米带MWCNTs/GONRs分散液的制备；步骤2、绿色碲化镉量子点溶液和红色碲化镉量子点溶液的制备；步骤3、碲化镉量子点?捕获探针分散液的制备：取步骤2制备的绿色碲化镉量子点溶液和红色碲化镉量子点溶液超声分散，采用HCl调节绿色碲化镉量子点溶液的pH值，得到混合液A；采用HCl调节红色碲化镉量子点溶液的pH值，得到混合液B；向混合液A中加入1?(3?二甲氨基丙基)?3?乙基碳二亚胺盐酸盐溶液，振荡均匀，加入N?羟基琥珀酰亚胺溶液，振荡均匀，加入终止子的探针，在室温下反应，得到绿色碲化镉量子点?捕获探针，记作NP?gQDs；向混合液B中加入1?(3?二甲氨基丙基)?3?乙基碳二亚胺盐酸盐溶液，振荡均匀，加入N?羟基琥珀酰亚胺溶液，振荡均匀，加入启动
 
申请号：CN201610714952.9
专利号：CN106404728-A
公开/公告号：CN106404728-A
申请/专利权人：江苏大学
发明/设计人：李雅琪;王坤;蔡健荣;孙力
分类号：G01N21/64
主分类号：G01N21/64
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