本文選題：楊梅　切入點：過敏原　出處：《浙江大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

【摘要】：對于大多數(shù)人來講,水果是一種健康食品,但有一部分人由于自身遺傳以及生活環(huán)境等因素,對某一種或者多種水果過敏。這種現(xiàn)象伴隨水果生產(chǎn)專業(yè)化以及銷售網(wǎng)絡(luò)全球化以及消費水果種類和數(shù)量的增加而更為普遍。水果過敏是患者的免疫系統(tǒng)發(fā)生紊亂而導(dǎo)致的,它主要是由于某些蛋白成分(過敏原)所引起的,一般屬于Ⅰ型超敏反應(yīng)(type I hypersensitivity),約1-3%的人群受到此類型疾病所困擾。因此,開展水果的過敏原研究對保護人們的生命健康、提高人民生活質(zhì)量具有巨大的經(jīng)濟效益以及社會效益。本論文主要是從楊梅過敏原蛋白的分離純化以及純化產(chǎn)物的免疫學(xué)活性分析等方面進行研究,得到如下結(jié)論： 首先,應(yīng)用Coca's提取法提取楊梅果實中粗蛋白液,運用還原SDS-PAGE技術(shù)對楊梅粗提蛋白液進行初步分析,顯示楊梅粗提蛋白中包含以前用過敏血清免疫印跡實驗鑒定為28kDa的主要過敏原蛋白條帶。 其次,通過超濾離心法、SP-FF離子交換層析以及Superdex7510/300GL凝膠柱層析方法分別純化楊梅主要過敏蛋白。依據(jù)楊梅轉(zhuǎn)錄組推導(dǎo)的氨基酸數(shù)據(jù),對楊梅主要過敏蛋白進行肽段指紋圖譜、分子量質(zhì)譜測定以及酶聯(lián)免疫(ELISA)間接法分析與鑒定,得到楊梅主要過敏原理論分子量和質(zhì)譜檢測所得分子量一致,ELISA檢測后有很強的免疫活性,且肽段指紋圖譜匹配實驗與氨基酸序列比對得出楊梅主要過敏蛋白屬于幾丁質(zhì)酶第三家族(class Ⅲ chitinase)(已提交至NCBI數(shù)據(jù)庫,序列號：KP307670)。 本研究初步對楊梅主要過敏原進行分離純化以及免疫活性的鑒定分析,可在ELISA檢測中作為楊梅特異性過敏原,為楊梅過敏反應(yīng)疾病的診斷奠定了良好的基礎(chǔ)。
[Abstract]:For most people, fruit is a healthy food, but some people because of their own inheritance and living environment and other factors, Allergies to one or more fruits are more common with specialization in fruit production, globalization of distribution networks, and an increase in the variety and quantity of fruit consumed. Fruit allergies are the result of a disorder in the patient's immune system. It is mainly caused by certain protein components (allergens), which generally belong to type I hypersensitivity, and about 1 to 3% of the population are affected by this type of disease. Therefore, the study of allergen in fruits can protect people's life and health. Improving people's quality of life has great economic and social benefits. In this paper, the purification of myricetin and the analysis of immunological activity of the purified product are studied, and the conclusions are as follows:. Firstly, the crude protein solution was extracted from the fruit of Myrica rubra by Coca's extraction method, and the crude protein extract of Myrica rubra was preliminarily analyzed by reducing SDS-PAGE technique. The results showed that the crude extract protein of Myrica rubra contained the main allergen protein bands identified as 28kDa by the immunoblotting assay of anaphylaxis serum. Secondly, the main allergic proteins of Myrica rubra were purified by ultrafiltration centrifugation with SP-FF ion exchange chromatography and Superdex7510/300GL gel column chromatography. According to the amino acid data derived from the transcriptome of Myrica myrica, the peptide fingerprint of the main allergic proteins of Myrica rubra was carried out. Molecular weight mass spectrometry and Elisa indirect analysis and identification showed that the theoretical molecular weight of the main allergen of Myrica rubra and the molecular weight of the main allergen detected by mass spectrometry were consistent with those of Elisa, and that the molecular weight of the main allergen of Myrica rubra had strong immunological activity. The peptide fingerprint matching experiment and amino acid sequence alignment showed that the main allergic protein of Myrica rubra belongs to the third family of chitinase (submitted to NCBI database, serial number: KP307670). In this study, the main allergens of Myrica rubra were isolated and purified and their immunological activities were identified and analyzed, which can be used as a specific allergen in the detection of Myrica rubra by ELISA, which lays a good foundation for the diagnosis of myrica anaphylaxis disease.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2015
【分類號】：R446.6

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