1. 研究目的与意义
CONTENT: 1. Fractionation of Rana chensinensis skin secretion by reverse phase HPLC. 2. Analysis of the sequence of the active natural peptide by LCQ. SIGNIFICANCE:To find novel bioactive peptides, identify their structures and provide new drugs for treatment of diseases.
2. 文献综述
Research advance on bioactive peptides from the skin extracts and secretions of the amphibian and its related technologyXiaowei FU(School of Pharmacy, Traditional Chinese Medicine, Nanjing University of Chinese Medicine, Nanjing, Jiangsu 210046, China)Abstract: The skin extracts and secretions of amphibian animals contains many kinds of bioactive peptides. Bioactive peptides have the functions which can regulate human physiological activities and metabolism. They are easy to be digested and absorbed by human bodies and play an important role in promoting immunity, regulating hormone, antibacterial, antiviral, lowering blood pressure and blood lipids and so on. However, up to now, for the reasons that the amphibian animal species are multitudinous and we are lack of systematic methods for the study of each species, the research and use pattern of bioactive peptides in amphibian animal skin are extensive, generally speaking, which cause a great waste of resources. In this paper, a series of achievements in the study of amphibian skin peptides which have been made in recent years are collected, inducted, reorganized and analyzed, such as methods of classifications, functions, separations, purification and structural identifications of the bioactive peptides from the skin extracts and secretions of the amphibian, which aims at discovering a large number of bioactive peptides, realizing the latest research technology about them, improving the utilization rate of bioactive peptide resourses from the skin extracts and secretions of amphibian animals and providing the basis for the discovery of novel antibiotics and anticancer drugs.Key words: amphibian; bioactive peptides; classification; functions; separation and purification; structural identification摘要：两栖类动物皮肤含有多种生物活性肽，在人体中生物活性肽能够调节新陈代谢和生理活动，并且易消化吸收，具有增强免疫力、调解激素分泌、抗菌、抗病毒、降血压、降血脂等作用。
然而目前为止，由于两栖类动物物种繁多，而我们还没有找到系统地针对这些物种进行研究的方法，因此我们对两栖类动物皮肤中生物活性肽的研发与利用方式，总体上还是粗放式的，这就造成了两栖类资源的浪费。
本文针对近年来在两栖类皮肤活性肽研究中取得的一系列成就做一归纳整理，例如生物活性肽的分类、功能、分离纯化以及结构鉴定，旨在系统发现大量生物活性多肽及以及与其相关的最新研究技术，提高两栖类动物的皮肤活性肽的资源利用率，并为新型抗生素和抗肿瘤药物的发现提供基础。
3. 设计方案和技术路线
RESEARCH PROGRAMME:1.Obtainment of skin secretion:Mince the skin of Chinese brown frog and dissolve it into 0.05%TFA.2.Analysis of skin peptides by RP-HPLC:Buffer A: purified water(contain 0.05% TFA).Buffer B: 80% acetonitrile(ACN, contain 0.05% TFA)Column: C-18 columns(250*10.00mm) Gradient elute: 100% A to 100% B in 240min Inject Volume:1000μL, Flow Rate: 1ml/min UV detection wavelength:214nm 280nm3.Analysis of skin peptides by LCQ: Method: Data Dependent Analysis (DDA) Solution: ACN(contain FA) Column: C-18 columns(100*4.60mm) NCE:30, Collision Mode:CID Inject Volume:20μL, Flow Rate 0.2ml/min ESI: Spray voltage: 4.5KV, Shealth gas:30, Aux gas: 10 Scan Range:300~2000 m/z 4.Database search and bioinformations analysis of peptidesS/N Threshold: 3.00, Database Uniprot_Amphibian, Precursor Mass Tolerance: 500ppm, Fragment Mass Tolerance:0.8Da, Dynamic Side Chain Modification: DeamidatedOxidationPhospho, Dynamic Peptide Modification: C-Terminus Amidated技术路线：见开题报告附件
4. 工作计划
01/03/2022~10/03/2022: Reading articles and writing review 10/03/2022~12/03/2022: Preparing the skin and cutting into powder14/03/2022~16/03/2022: Extracting and purifying the skin peptides17/03/2022~21/03/2022:Analyzing by RP-HPLC22/03/2022~25/03/2022: Analyzing by LCQ25/03/2022~31/03/2022: Database search01/04/2022~20/05/2022: Write the paper
5. 难点与创新点
We first find all categories of peptides in the skin of Rana chensinensis, which contains the novel peptides that have not been identified yet. This work will finally provide new methods or ideas for the development of antimicrobial drugs by finding novel bioactive peptides.
以上是毕业论文开题报告，课题毕业论文、任务书、外文翻译、程序设计、图纸设计等资料可联系客服协助查找。
