基于石墨烯氧化物-DNA傳感器對汞離子高靈敏檢測的研究
發(fā)布時間:2018-04-18 08:00
本文選題:石墨烯氧化物 + 汞離子; 參考:《江蘇大學(xué)》2017年碩士論文
【摘要】:汞離子(Hg~(2+))是環(huán)境污染中的重要因素之一,由于該物質(zhì)低濃度即可對動植物帶來不可逆的影響,經(jīng)過循環(huán)過程離子濃度被不斷富集,對人類的身體健康造成不可估量的傷害。Hg~(2+)可以與特異性核酸序列,如多T的DNA序列作用,引起DNA構(gòu)型發(fā)生變化,基于這個原理一些檢測方法被發(fā)展,但仍然存在的一些問題:有的方法檢測樣本少、有的假陽性信號率高、有的靈敏性較低等,為解決這些問題,將Hg~(2+)特異作用的DNA作為識別元件結(jié)合石墨烯氧化物(GO)構(gòu)建生物傳感器。生物傳感器是分子生物學(xué)的一種研究工具,基于生物傳感器對Hg~(2+)進(jìn)行檢測,發(fā)展檢測的新方法與新技術(shù),進(jìn)行以下幾個方面的研究:1.GO表面固定多T序列對汞離子的檢測。多T的DNA序列作為識別元件,羧基熒光素(FAM)修飾的DNA序列固定到作為淬滅劑的GO表面,去除沒有結(jié)合上的DNA,減少了假陽性對檢測的影響,提高檢測的靈敏性,檢測極限為9.37 nM,比物理吸附降低了20%。進(jìn)一步通過與檢測序列互補的DNA序列進(jìn)行雙氨基固定,雙氨基修飾的DNA檢測極限為2.52 nM,降低了75%。2.雙熒光標(biāo)記多T序列對汞離子的檢測。用FAM和SBRY Green I兩種熒光染料對DNA序列標(biāo)記,這兩種熒光基團協(xié)同作用,相互促進(jìn)。單氨基修飾的DNA序列結(jié)合雙熒光修飾構(gòu)建的傳感器檢測極限為0.53nM,雙氨基修飾的DNA檢測極限為0.43 nM,降低了20%。3.硫代磷酸化(PS)修飾的DNA序列固定到GO表面對汞離子檢測。Hg~(2+)能特異性切割PS修飾的DNA,使Hg~(2+)能夠重復(fù)使用,用FAM標(biāo)記的PS修飾的DNA固定在GO表面對Hg~(2+)檢測的極限為1.37 nM,比物理吸附法降低了22%。4.硫代磷酸化修飾的DNA序列結(jié)合生物芯片對汞離子的檢測。將多個DNA序列樣品在玻片上進(jìn)行點陣排列,加入Hg~(2+)實現(xiàn)對多樣品的檢測。GO表面物理吸附PS修飾的DNA序列的檢測極限為3.187 nM,固定PS修飾的DNA序列的檢測極限為0.79 nM。通過上述研究,解決現(xiàn)有Hg~(2+)檢測方法中存在的問題,發(fā)展了幾種Hg~(2+)檢測的新方法與新技術(shù),推動食品安全中重金屬檢測技術(shù)的進(jìn)一步發(fā)展。
[Abstract]:Mercury ion (HG) is one of the important factors in environmental pollution. Because the low concentration of the substance can bring irreversible effects on animals and plants, the ion concentration is continuously enriched through the cycle process.Can act with specific nucleic acid sequences, such as polyT DNA sequences, causing changes in the configuration of DNA. Based on this principle, some detection methods have been developed.But there are still some problems: some methods to detect small samples, some false positive signal rate, some low sensitivity, to solve these problems,The biosensor was constructed by using Hg~(2 (DNA) as recognition element and graphene oxide (GOG).Biosensor is a research tool in molecular biology. The detection of Hg~(2 based on biosensor and the development of new methods and techniques are studied in the following aspects: 1. The detection of mercury ions by multi-T sequences fixed on the surface of go.The polyT DNA sequence was used as the recognition element, and the carboxyfluorescein modified DNA sequence was immobilized to the surface of go as a quenching agent to remove the unbound DNA, thus reducing the influence of false positive on the detection and improving the sensitivity of the detection.The detection limit is 9.37 nm, which is 20% lower than physical adsorption.Further, the DNA sequence complementary to the detection sequence was immobilized with two amino groups. The detection limit of the DNA modified by the double amino group was 2.52 nm, which reduced the detection limit by 75. 2.Detection of mercury ions by double fluorescence labeling multiple T sequences.The FAM and SBRY Green I fluorescent dyes were used to label the DNA sequence. The two fluorescent groups acted synergistically and promoted each other.The detection limit of single amino modified DNA sequence combined with double fluorescence modification was 0.53 nm and the detection limit of double amino modified DNA was 0.43 nm, which decreased 20.3 nm.Thiophosphorylated DNA sequence immobilized on go surface to detect mercury ions. Hgf2) can specifically cleave PS modified DNA so that Hg~(2) can be reused.The detection limit of DNA modified with PS modified by FAM on go surface is 1.37 nm, which is 22. 4% lower than that of physical adsorption method.Thiophosphorylation modified DNA sequence combined with biochip to detect mercury ions.The detection limit of PS modified DNA sequence is 3.187 nm and the detection limit of fixed PS modified DNA sequence is 0.79 nm.Through the above research, the problems existing in the existing Hg~(2) detection methods are solved, and several new methods and technologies of Hg~(2) detection are developed to promote the further development of heavy metal detection technology in food safety.
【學(xué)位授予單位】:江蘇大學(xué)
【學(xué)位級別】:碩士
【學(xué)位授予年份】:2017
【分類號】:Q503;TP212
【參考文獻(xiàn)】
相關(guān)期刊論文 前2條
1 趙秋伶;張尤華;于曉艷;;基于核酸外切酶Ⅰ的適配體熒光傳感器檢測人尿液中的Hg~(2+)[J];分析化學(xué);2016年07期
2 莫志宏;楊琳玲;楊小超;陳自鋒;;基于胸腺嘧啶-汞離子-胸腺嘧啶結(jié)構(gòu)和納米金放大傳感器檢測汞離子[J];分析化學(xué);2009年07期
,本文編號:1767488
本文鏈接:http://sikaile.net/shoufeilunwen/xixikjs/1767488.html
最近更新
教材專著