本文選題：金納米粒子　切入點(diǎn)：核酸適配體　出處：《河北大學(xué)》2008年碩士論文

【摘要】： 蛋白質(zhì)是構(gòu)成生物體的重要組成部分之一,是一切生命的物質(zhì)基礎(chǔ),承載著生物體完成各項(xiàng)生物功能的任務(wù)。因此,蛋白質(zhì)的分析在生命科學(xué)的研究中具有重要的意義,廣泛應(yīng)用于生物學(xué)、醫(yī)學(xué)診斷等領(lǐng)域。傳統(tǒng)的蛋白質(zhì)檢測(cè)往往需要熒光標(biāo)記、凝膠電泳和放射性自顯影等繁雜操作,在一定程度上限制了人們對(duì)這些生命物質(zhì)的深入研究。隨著生命科學(xué)的發(fā)展,如何對(duì)蛋白質(zhì)進(jìn)行快速、簡(jiǎn)便、高效分析,已經(jīng)成為研究的熱點(diǎn)問(wèn)題。 本文以凝血酶為研究模型,利用核酸適配體(aptamer)與金納米粒子(gold nanoparticle)技術(shù)相結(jié)合制作生物傳感器,以核酸適配體作為蛋白質(zhì)識(shí)別元件,發(fā)展了蛋白質(zhì)分析的新方法,該方法快速、靈敏、操作簡(jiǎn)單而且具有很高的選擇性。本文分為兩個(gè)部分： 第一部分：基于核酸適配體標(biāo)記的金納米粒子探針測(cè)定凝血酶的共振光散射分析。 核酸適配體是一種具有特殊功能的寡聚核苷酸,它能與目標(biāo)分子,包括核酸、蛋白質(zhì)、藥物等特異性結(jié)合,從而實(shí)現(xiàn)目標(biāo)分子的測(cè)定。以標(biāo)記核酸適配體的金納米粒子為探針,通過(guò)核酸適配體與凝血酶的特異性作用,金納米粒子發(fā)生自組裝,形成網(wǎng)狀結(jié)構(gòu),導(dǎo)致共振光散射信號(hào)的增強(qiáng),從而定量測(cè)定了a凝血酶。該方法操作簡(jiǎn)便,具有較高的靈敏度,為蛋白質(zhì)的檢測(cè)開(kāi)辟了一條新途徑。 第二部分：基于匹配DNA鏈標(biāo)記的金納米粒子探針與核酸適配體作用測(cè)定凝血酶的共振光散射分析。 本文設(shè)計(jì)了兩條與核酸適配體匹配的寡核苷酸鏈并將其分別標(biāo)記到金納米粒子上,當(dāng)金納米粒子探針與核酸適配體混合,此探針與核酸適配體的雜交導(dǎo)致金納米粒子的聚集并引起共振光散射的強(qiáng)烈增強(qiáng)；而核酸適配體與凝血酶的特異性結(jié)合將抑制此探針與核酸適配體的雜交并導(dǎo)致共振光散射強(qiáng)度的降低,降低的程度與凝血酶的濃度有關(guān),從而可以定量測(cè)定凝血酶。該方法只要改變核酸適配體和相應(yīng)的探針序列,即可有望用于其它蛋白質(zhì)的分析,為蛋白質(zhì)的檢測(cè)提供了新的思路和手段。
[Abstract]:Protein is one of the important components of organism and the material basis of all life.Therefore, protein analysis plays an important role in life science research and is widely used in biology, medical diagnosis and other fields.Traditional protein detection often requires a lot of operations such as fluorescent labeling, gel electrophoresis and radioautography, which to some extent limits the further study of these living substances.With the development of life science, how to analyze proteins quickly, easily and efficiently has become a hot issue.In this paper, a novel method for protein analysis was developed by using thrombin as a research model, using aptamer (a nucleic acid aptamer) and gold nanoparticles (gold nanoparticles articlein) technology to produce biosensor, using aptamer of nucleic acid as a protein recognition element.It is sensitive, simple and selective.This paper is divided into two parts:Part I: resonance light scattering (RLS) analysis of thrombin by gold nanoparticles probe labeled with aptamer of nucleic acid.Nucleic acid aptamer is a kind of oligonucleotide with special function. It can bind to target molecule, including nucleic acid, protein, drug and so on, so as to realize the determination of target molecule.Gold nanoparticles labeled with nucleic acid aptamers were used as probes. Through the specific interaction between aptamers and thrombin, gold nanoparticles were self-assembled and formed a network structure, which led to the enhancement of resonance light scattering (RLS) signals.Thus, a thrombin was quantitatively determined.The method is easy to operate and has high sensitivity, which opens a new way for protein detection.Part two: resonance light scattering (RLS) analysis of thrombin based on the interaction between gold nanoparticles labeled with DNA chain and aptamer of nucleic acid.Two oligonucleotide chains matching the aptamer of nucleic acid were designed and labeled onto gold nanoparticles respectively when the gold nanoparticles probe was mixed with the aptamer of nucleic acid.The hybridization of the probe with the aptamer of nucleic acid leads to the aggregation of gold nanoparticles and the enhancement of resonance light scattering.The specific binding of aptamer and thrombin will inhibit the hybridization between the probe and the aptamer of nucleic acid and lead to the decrease of the intensity of resonance light scattering (RLS), which is related to the concentration of thrombin, so the thrombin can be quantitatively determined.This method can be used in the analysis of other proteins by changing the aptamer of nucleic acid and the corresponding probe sequence, which provides a new way for protein detection.
【學(xué)位授予單位】：河北大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R346

【引證文獻(xiàn)】

相關(guān)期刊論文 前1條

1 李瑜;王源升;于斌;張靜秋;董瑞;;核酸適配體在光生化檢測(cè)中的應(yīng)用[J];廣州化工;2012年14期

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本文編號(hào)：1709782