黄芩诱导型Bax基因工程细胞株的构建-食品科学专业毕业论文.docxVIP

黄芩诱导型Bax基因工程细胞株的构建20mg／L的培养基上选择培养30天后，获得了转化细胞。经过PCR 黄芩诱导型Bax基因工程细胞株的构建 20mg／L的培养基上选择培养30天后，获得了转化细胞。经过PCR 及Western blotting检测后证明Bax已经整合到了黄芩染色体基因 组上，并且在诱导剂雌二醇存在的情况下能够稳定表达。随后我们利 用得到的转基因细胞建立了转基因悬浮细胞系。为了观察Bax基因调 控下黄芩细胞凋亡情况，又分别将转基因黄芩细胞和野生型细胞置于 雌二醇浓度50 p M的培养基上培养3天后提取DNA，电泳后观察到了 转基因黄芩细胞DNA出现梯带降解情况。细胞核形态学观察同时表 明，经雌二醇处理的转基因黄芩悬浮细胞细胞核出现皱缩、细胞膜破 裂等凋亡特征。试验结果证实，小鼠Bax基因不仅可以在黄芩细胞中 稳定表达并且能够触发黄芩细胞凋亡。本研究获得了稳定的黄芩诱导 型Bax基因工程细胞系，为进一步研究探讨Bax基因对黄芩细胞次生 代谢的调控作用及分子机制提供了必不可少的实验材料。有关Bax基 因对转基因黄芩细胞次生代谢的调控及其分子机制等方面的研究工 作目前正在进行中。 关键词黄芩，次生代谢，Bax基因，雌二醇，细胞凋亡 II 黄芩诱导型Bax基因工程细胞株的构建CoNSTRUCTIoN 黄芩诱导型Bax基因工程细胞株的构建 CoNSTRUCTIoN oF INDUCIBLE BAX TRANSGENIC CELLS OF Scutellaria baicalensis Georgi Abstract Hi。gher plants constitute one of our most important natural resources，which provide not only foodstuffs，fibers，and woods，but also many chemicals，such as flavorings，dyes，and pharmaceuticals． Although plants are renewable resources，some species are becoming more difficult to obtain in sufficient amounts to meet increasing demands．Plant cell cultures ofrer promising alternatives for the production of the useful secondary metabolites because totipotency enables plant cells to produce the chemicals in vitro．After extensive empirical trials，some metabolites are now being produced by large—scale cell culture，e g shikonin and berberine，but the numbers of compounds that are producible commercially by cell culture technology are still very few．One of the main limitations is the IOW productivity of the desired compounds．Molecular engineering of secondary metabolites has the potential to increase productivity and improve product composition． Apoptosis(or programmed cell death，PCD)is a suicide procedure regulated strictly by genes in plants and animals．In plants，Hypersensitive Reaction(HR)is activated during PCD， and the system resistance is obtained by the activation of some second metabolic pathways，which implies that the secondary metabolite productivity of plant cells could be ireproved by triggering PCD．Bax is an