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低剪切力模拟微重力环境对巴氏毕赤酵母表达重组人源胶原蛋白的生物学效应

时间:2017-07-27 20:49来源:毕业论文
利用高纵比培养容器模拟空间微重力环境,考察在微重力条件下重组基因工程菌对表达胶原蛋白的影响,利用二种不同重力条件(低剪切力模拟微重力、常规重力)和不同时间(24h, 48h,
胶原蛋白是广泛存在于低等脊椎动物和哺乳动物机体中的一类蛋白质,由于其在提高免疫机能、抑制癌细胞和活化细胞机能等方面具有许多优势特性,在食品、制药工业等诸多领域有广泛的应用。
重力在生物进化中,扮演着十分重要的角色。而模拟微重力环境对微生物和细胞有特殊的生物学效应。
本毕业设计利用高纵比培养容器模拟空间微重力环境,考察在微重力条件下重组基因工程菌对表达胶原蛋白的影响。
本实验利用二种不同重力条件(低剪切力模拟微重力、常规重力)和不同时间(24h, 48h, 72h, 96h, 120h,144h,168h, 192h, 216h)条件下培养的基因重组工程菌,在低剪切力模拟微重力条件下对表达重组类人胶原蛋白量较高的基因重组工程菌进行重复试验,初步试验结果表明:低剪切力模拟微重力试验条件下培养的基因重组工程菌,菌体湿重与对照组比较,提高了46%。同时利用凝胶柱过滤层析法分离纯化发酵液,得到的纯化产物通过SDS-PAGE电泳、相对分子质量测试进行表征,并与前期本实验室获得的重组类人胶原蛋白的标样谱图比对,其图谱所提供的信息显示:本实验中重组毕赤酵母所表达的重组类人胶原蛋白与前期发酵罐获得的类人胶原蛋白具有分子质量上的一致性。与摇瓶相比产量有明显提高。12206
关键词:  重组毕赤酵母 空间微重力条件 重组类人胶原蛋白 低剪切力
毕业设计说明书(论文)外文摘要

Title The effects of low shear modeled microgravity (LSMMG) on the characterization of recombinant Human like collagen expressed in Pichia pastoris
Abstract
Collagen is a series of protein which is widely Presenting in the lower vertebrates and mammals. Collagen has increasing been uesd in various fields, such as food industry, medical and health industry, etc., because of its special effects on improving immune function, restraining cancer cell and activating cell function.

源自六/维"论\文|网.加7位QQ3249"114 www.lwfree.cn



Gravity plays an important role in evolution of life on Earth and provides the required living environment for life activities. Microgravity have special effect on microbes and cells.
We try to research on different effects caused by different microgravity conditions, by using ground based high-aspect-ratio vessel (HARV) to model microgravity .

Two groups of samples, which are handled with low shear modeled microgravity (LSMMG) and normal gravity(NG) 24h, 48h, 72h, 96h, 120h, 144h, 168h have been tested, respectively. After three rounds of fermentation on recombinant Human like collagen expressed in Pichia pastori, we could draw the conclusion from the results that Pichia pastoris cultured in LSMMG have higher production which output of recombinant human-like collagen increased 46% when compared with Pichia pastoris cultured in NG.

At the moment, product was obtained through purifying the fermentation supernatant by gel filtration chromatography. Comparing the results of SDS-PAGE Gel electrophoresis of human-like collagen and previous target protein we obtained, we presume that purified recombinant human-like collagen is similar to animal-derived gelatin in protein and molecular mass.
Keywords: recombinant Pichia pastoris; Human like collagen; low shear modeled microgravity (LSMMG);high-aspect-ratio vessel (HARV)
目次
1 绪论    2
1.1胶原蛋白简介    2
1.1.1胶原蛋白的结构    2
1.1.2  胶原蛋白的类型    3
1.1.3  胶原蛋白的生产    3
1.1.4  胶原蛋白的功能    4
1.2 低剪切力模拟微重力环境对微生物生长的影响    5
1.3 毕赤酵母表达人源胶原蛋白简介    6
1.4 人源胶原蛋白的分离纯化    6 低剪切力模拟微重力环境对巴氏毕赤酵母表达重组人源胶原蛋白的生物学效应:http://www.lwfree.cn/shengwu/20170727/11902.html
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