本文選題：非病毒基因載體 + 魚(yú)精蛋白��； 參考：《上海交通大學(xué)》2007年碩士論文

【摘要】： 本文研究了硬脂酰魚(yú)精蛋白的合成,并將其作為非病毒基因載體,考察了與質(zhì)粒DNA的復(fù)合性能,復(fù)合物的體外基因轉(zhuǎn)染效果。主要內(nèi)容包括以下三部分: 1)通過(guò)N-羥基琥珀酰亞胺(NHS)與硬脂酸形成的活潑酯,在水性反應(yīng)環(huán)境中與魚(yú)精蛋白進(jìn)行酯交換反應(yīng),從而將疏水性的硬脂�；鶊F(tuán)偶聯(lián)到魚(yú)精蛋白上,成功合成了三種硬脂酰魚(yú)精蛋白(stearyl protamine,簡(jiǎn)稱SP),分別為SP1,SP2,SP3。將SP在HCl中降解后回收硬脂酸,并用酸堿微量滴定的方法測(cè)定含量。SP中硬脂�；鶊F(tuán)的含量(質(zhì)量百分比)分別為2.5%、3.0%、5.5%。通過(guò)動(dòng)態(tài)光散射(DLS)對(duì)SP以及硫酸魚(yú)精蛋白(PS)的研究發(fā)現(xiàn),SP3可以在水中自組裝形粒徑110nm左右的穩(wěn)定聚集體,此聚集體的粒徑不隨SP3濃度的提高而發(fā)生變化,說(shuō)明是一種熱力學(xué)穩(wěn)定的自組裝聚集體。 2)通過(guò)工程菌擴(kuò)增,堿法抽提,獲得編碼螢火蟲(chóng)熒光素酶的質(zhì)粒DNA pGL3-control,并以此作為后繼實(shí)驗(yàn)中采用的DNA材料。凝膠電泳阻滯實(shí)驗(yàn)檢驗(yàn)SP對(duì)DNA的復(fù)合能力,結(jié)果表明當(dāng)載體/DNA質(zhì)量比超過(guò)1.0時(shí),DNA均能被三種SP完全包裝起來(lái)。與未修飾的魚(yú)精蛋白相比,隨著SP中硬脂�；鶊F(tuán)含量的升高,其對(duì)DNA的復(fù)合能力略有減弱。對(duì)魚(yú)
[Abstract]:In this paper, the synthesis of stearyl protamine was studied, and it was used as a non-viral gene vector. The compound with plasmid DNA and the effect of gene transfection in vitro were investigated. The main contents include the following three parts: 1) the hydrophobic stearyl group was coupled to protamine by transesterification of active ester formed by N-hydroxysuccinimide (NHS) with stearic acid. Three stearyl protamine (SPN) stearyl protamines were successfully synthesized. The stearic acid was recovered after the degradation of SP in HCl, and the content (mass percentage) of stearyl group in SP was determined by acid-base microtitration. The content of stearyl group (mass percentage) was 2.5% and 3.5% respectively. The study of SP and protamine sulfate PSs by dynamic light scattering (DLS) showed that SP3 could be used to form stable aggregates with self-assembled particle size (110nm) in water, and the particle size of the aggregates did not change with the increase of SP3 concentration. It shows that it is a thermodynamically stable self-assembly aggregate. 2) the plasmids DNA pGL3-control encoding luciferase of firefly were obtained by amplification and alkali extraction, which were used as DNA materials in subsequent experiments. The results of gel electrophoresis block test showed that when the mass ratio of vector / DNA was more than 1.0, the DNA could be completely packaged by three kinds of SP. Compared with unmodified protamine, the compound ability of SP to DNA decreased slightly with the increase of stearyl group content in SP. Yeah, fish.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R346

【共引文獻(xiàn)】

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