本文選題：銀納米簇　切入點：D-甘露糖　出處：《青島科技大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目前,具備熒光性質(zhì)的納米材料發(fā)展十分迅速,尤其是以DNA為模板合成的銀納米簇(Ag NCs),其最大的特點是可以通過變換不同的DNA序列和長度能夠?qū)崿F(xiàn)從可見光到近紅外的熒光可調(diào)控發(fā)射。此外,它還具有制備方法簡單、光穩(wěn)定性好、低毒性、生物相容性良好的優(yōu)點,因此受到很多研究學(xué)者的青睞。本論文對DNA穩(wěn)定的銀納米簇展開研究和應(yīng)用,包括利用糖修飾銀納米簇檢測伴刀豆蛋白A、腫瘤細(xì)胞成像檢測、利用銀納米簇與T-Hg~(2+)-T的復(fù)合結(jié)構(gòu)對汞離子進(jìn)行靈敏檢測,主要開展以下研究。一是,基于DNA穩(wěn)定的銀納米簇和D-甘露糖與伴刀豆蛋白A(Con A)的特異性結(jié)合作用,我們實現(xiàn)Con A的熒光檢測。設(shè)計方案是:N-DNA富含胞嘧啶,作為銀納米簇的成核序列,此時制備的銀納米簇?zé)晒馕⑷?G-DNA鏈富含鳥嘌呤,當(dāng)富含鳥嘌呤的G-DNA序列靠近銀納米簇時,其熒光強(qiáng)度會明顯增強(qiáng)。D-甘露糖修飾在兩條DNA鏈的不同末端。當(dāng)有Con A存在時,就會誘導(dǎo)兩條DNA鏈靠近,從而使熒光增強(qiáng),達(dá)到檢測Con A的目的。Con A在1.0×10-12到1.0×10-7M之間時,其濃度與體系的熒光強(qiáng)度成良好的線性關(guān)系。二是,基于DNA穩(wěn)定的銀納米簇和D-甘露糖與MDA-MB-231細(xì)胞表面特異性受體識別的作用,通過成像手段檢測腫瘤細(xì)胞。實驗首先用巰基乙酸糖化制備羧基-α-D-甘露糖,通過羧基與氨基的共價作用,使得甘露糖修飾在DNA序列的末端,該DNA帶有銀納米簇的成核序列。當(dāng)DNA與硝酸銀和硼氫化鈉作用之后就會形成銀納米簇。當(dāng)與細(xì)胞一起培育時,該熒光探針就會與細(xì)胞表面受體特異性結(jié)合,從而實現(xiàn)癌細(xì)胞識別成像研究。三是,由于DNA與Hg~(2+)有特殊的相互作用,能夠形成T-Hg~(2+)-T的復(fù)合結(jié)構(gòu),我們開發(fā)出一種免標(biāo)記檢測汞離子的簡便方法。實驗中我們設(shè)計一個汞離子存在時能通過T-Hg~(2+)-T結(jié)構(gòu)形成莖環(huán)結(jié)構(gòu)的DNA序列。加入Exo III對莖環(huán)結(jié)構(gòu)進(jìn)行剪切,釋放出目標(biāo)物Hg~(2+)和銀納米簇的模板鏈,釋放出的目標(biāo)物Hg~(2+)能夠再次進(jìn)入到循環(huán)中,形成新的莖環(huán)結(jié)構(gòu)。利用這種方法,我們實現(xiàn)了Hg~(2+)的定量檢測,Hg~(2+)的濃度在3.0×10-11到3.0×10-8 M時,濃度與熒光強(qiáng)度呈良好的線性關(guān)系。
[Abstract]:At present, with the development of the fluorescence properties of nano materials is very rapid, especially with DNA as the template for the synthesis of silver nanoclusters (Ag NCs), its biggest feature is through the DNA sequence and length transform can realize different from visible to near infrared fluorescence emission can be controlled. In addition, it also has the preparation the method is simple, good light stability, low toxicity, good biocompatibility and good, so by many scholars of all ages. This thesis research and application of silver nanoclusters DNA stable, including the detection of the use of sugar modified silver nanoclusters with egg white concanavalin A, tumor cell imaging detection, the use of silver nanoclusters and T-Hg~ (2+) -T composite structure for sensitive detection of mercury ions, mainly in the following research. First, the DNA stability of silver nanoclusters and D- mannose with concanavalin A (Con A) based on the specific binding, we implement Con A fluorescence detection Test. The design scheme is: N-DNA is rich in cytosine, as nuclear sequence into silver nanoclusters, fluorescent silver nanoclusters prepared at the weak, G-DNA chain guanine rich guanine rich sequences of G-DNA, when close to the silver nanoclusters, the fluorescence intensity will be significantly enhanced.D- mannose modified in different end of two strands of DNA. When Con A, will be induced by two DNA strands near, so that the fluorescence enhancement, to detect Con A to.Con A in the 1 * 10-12 to 1 * 10-7M, the fluorescence intensity of the concentration and the system of a good linear relationship. The two is that the DNA stability of silver nanoclusters and D- mannose with the specific MDA-MB-231 cell surface receptor recognition based on the effect of tumor cell detection by imaging means. The first experiment with mercaptoacetic acid saccharification and preparation of carboxylated alpha -D- mannose, through covalent interactions of carboxyl and amino groups, the mannose modification in DNA sequence In the end, the DNA sequence of nucleation with silver nanoclusters. After DNA with silver nitrate and sodium borohydride will form silver nanoclusters. When cultivated together with cells, combined with the fluorescent probe will with a cell surface receptor specificity, so as to realize the imaging of cancer cell recognition. The three is due to DNA and Hg~ (2+) has special interaction, can form T-Hg~ (2+) -T composite structure, we developed a simple method for label free detection of mercury ions. In our experiment design through the T-Hg~ there is a mercury ion (2+) -T formation sequence of DNA stem loop structure. Adding Exo III cut the stem loop structure, release the object Hg~ (2+) and silver nanoclusters template chain, the release of the target Hg~ (2+) to enter into circulation again, form a stem loop structure. Using this new method, we implement the Hg~ (2+) quantitative detection, Hg~ (2+ concentrated) When the degree is 3 * 10-11 to 3 x 10-8 M, the concentration has a good linear relationship with the fluorescence intensity.

【學(xué)位授予單位】：青島科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：O657.3

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 汪舒婷;張權(quán);葉舟;熊永權(quán);崔晨宇;尹健;;D-甘露糖修飾聚合物膠束的制備及其在靶向藥物輸送中的應(yīng)用[J];生物工程學(xué)報;2016年01期

2 周國強(qiáng);陳春英;李玉鋒;李煒;高愈希;趙宇亮;;納米材料生物效應(yīng)研究進(jìn)展[J];生物化學(xué)與生物物理進(jìn)展;2008年09期

3 鮑錦庫,曾仲奎,周紅,楊莉;巖豆凝集素分子修飾與圓二色性的關(guān)系研究[J];中國生物化學(xué)與分子生物學(xué)報;1998年06期

，

本文編號：1558732