《基于发卡DNA自组装金胶比色检测DNA及分子识别文库的构建》由会员分享，可在线阅读，更多相关《基于发卡DNA自组装金胶比色检测DNA及分子识别文库的构建（87页珍藏版）》请在金锄头文库上搜索。
1、基于发卡DNA皂塑茎全壁些鱼垡型里竖垒垄坌王塑型茎壁塑塑堡_-一片段之间同源性较低组合变化较多，达到实验的预期目标。此方法的创建必将对难以找到识别分子的检测目标如新的靶点受体的筛选，预防和治疗新发现的病毒等发挥积极的作用。关键词：纳米金信号放大DNA分子识别文库重聚PCR青岛科技大学研究生学位论文CoLoRIMETRIC DNA DETECTIoN BASED oNHAIRPIN ASSEMBLY REACTIoN ANDCoNSTRUCTIoN oF RECoGNIZING MoLECULARGENE LIBRARYABSTRACT1、Simple colorimetric DNA dete
2、ction based 011 hairpin assembl37 reaction andtargetcatalytic circuits for signal amplificationDNA is the genetic factor in organismsThe detection of DNA is often connectedto the diagnosis of different diseaseSO the detection of DNA is ver)7 meanillgful tostud7Because the progress of detection techn
3、ology for DNAhuman ha、e changedthe detection of other biological molecules to the detection of DNAfor exampleprotein and small molecular substancesThere are many methods of DNA detectionfor example：fluorescence detection methodchemi luminescence detectioncolorimetry detectionelectrochemical detectio
4、n and fluorescence quantitative PCRand SO onIn the method of biochemical analy,sisthe hot issue is increasin。,thesensitivity of detectionSO the technology of signal amplification is widely usedThetool enzymebased signal amplification and gold nanoparticle biological bar code isthe hot issue of study
5、Our topic is based 011 biological compatibilityof goldnanoparticle，binding DNA probe onto the gold nanoparticle to realize the signalamplificationand finally，detect the target DNA by gold nanoparticle colorimetryA simple strategy of colorimetric DNA detection was presented in the presentwork，which w
6、as based on hairpin assembly reaction and targetcatalytic DNA circuitsto realize enzymefree signal amplificationThe method employed two hairpin speciesH 1 and H2，hich were stable and unable to hybridize in the absence of targetIn thepresence of targetthe target hybridized with hairpin H 1 and the op
7、ened hairpin H 1hybridized with hairpin H2，which made the target be displacedThe duplex of hairpinH 1 and H2 modified on nanoparticles made particle aggregation be visualized with1基于发卡DNA自组装金胶比色检测DNA及分子识别文库的构建the naked eyesThe displaced target again triggered the next round ofstrandexchange reaction
8、 to realize signal amplificationThe method would have awide range of sensing applications7ith enzymefreeeasy operation and extendedfield of applicationOur approach is simple，sensitiveand selectiveThis approach could be appliedto a wide range of sensing applicationsithout enzymeeasy operation and ext
9、endedfield of applicationThe method could detect as little as 22nM target DNA and ishighl3selectixe for completely complementarDN AThe approach，ould al so be used todetect extensive targets including ploteinssmall moleculesand metal ions that callproduce initiator DNA strand111 additionthis approach
10、 may be a potentially usefultool in the fields ofbasic and clinical researchand diagnostics2、Analysis and design of structure domain and preliminary optimization ofrecombination for geneBiological molecular recognition element is an important part of biologicalsensolLooking for a Varietvof biologica
11、l molecular recognition element is one oftheimportant development directions of biological sensorIn this partwe mainly use thecombined gene modules of directed evolution methods to construct the diversity ofgene library，searching for novel protein recognition moleculeFor convenience of screeningthis
12、 experiment takes kanamycin resistance geneprotein as recognition molecules to verif7 the identification of library constructionmethod is effectiveTake the kanamycin resistance gene for example，we explored theprocess of directed evolutionFirst of a11，e looked up the kanamycin resistance genefrom Gen
13、eBank and analysis the gene sequenceAccording to the twodimensionalstructure we designed the DNA segment to carry through the reassembly PCRIn ourworkwe mostly optimized the condition of the reassembly PCRamplified thereassembl，v gene product，connected with the career and transformed into Ecolibacte
14、riaAt last。we randomly chose the proper length gene fragment and measuredthe sequenceThe result displayed poor homology between them as we expectedThecreating of this method will play a positive role for detection of tmget that is hard toidentify,for example：screening of newtarget receptor and preve
15、ntion and treatmentof the newly discovered virusKEY WORDS：Gold Nanoparticle Signal Amplification DNA MolecularRrecognition Eelement Llibrary Reassembly PCR目 录第一章基于发卡DNA自组装金胶比色检测DNA11前言111 DNA检贝01111 DNA检测的意义1112 DNA捡湖0的力法112信号放大的方式4121基于工具酶的信号放大技术5122纳米金信号放火技术8123熵驱动催化反应信号放_7重聚产物上游引物下游引物dNTP】0bufferPfuEM924H20反应程序为：1 uL04 uL04 uL14 uL2 uL01 uL12 uL补至20 uL青岛科技大学研宄生学位论文9452茹C 30 S)50cycles72C 1 rain。 。 J选取扩
