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所在平台: Udemy |
课程主页: https://www.udemy.com/course/biotechnology-basics-of-gene-cloning-and-gene-therapy/
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课程名称:生物技术:基因克隆与基因治疗基础 课程概述: 近年来,生物技术因基因克隆而备受关注。尽管通过微生物培养可以获得许多有用的产品,但过去仅限于微生物自然合成的化合物。许多重要药物是由高级生物体产生的,无法通过这种方式获得,而基因克隆的应用改变了这一现状。基因克隆涉及从较大的染色体中分离特定基因或DNA片段,将其附着到小分子的载体DNA上,然后通过增加宿主细胞数量和在每个细胞中创建多个克隆DNA副本的方式,重复这一修改后的DNA数千或数百万次。 基因克隆的重要环节是通过数十年的核酸代谢研究而获得的一系列酶。经典的重组DNA分子的生成和传播方法中,限制性核酸内切酶和DNA连接酶是核心酶。限制性内切酶能够识别并切割DNA特定序列,生成一组较小的片段。某些限制性内切酶在两条DNA链上产生错位切口,留下未配对的粘性端,而另一些则在磷酸二酯键上切割两条DNA链,形成平端。 重组DNA分子构造的最后一步是连接克隆载体和需要克隆的基因,这个过程称为连接,催化该反应的酶是DNA连接酶。克隆载体是能够在生物体中稳定维持的小DNA片段,外源DNA片段可以插入其中进行克隆。可用的克隆载体种类繁多,但选择合适的载体需考虑插入片段的大小、拷贝数和克隆方法等因素。一旦基因被克隆,可以获得关于其结构和表达几乎无限的信息。克隆材料的可得性促进了研究基因的分析方法的发展,新的技术不断被引入。 重组DNA技术在试图治愈遗传性疾病方面显示出了价值。基因治疗最初是指通过提供患者正确的缺陷基因副本来治愈遗传病的方法,现在已扩展到通过将克隆基因引入患者体内来治愈任何疾病。基因治疗的临床应用并不限于遗传病的治疗。大多数癌症因癌基因的激活导致肿瘤形成,或抑癌基因的失活而导致肿瘤形成。面对这两种情况,都可以设想通过基因治疗来治疗癌症。 本课程是生物学、法医科学、分子生物学、生物技术、生物化学和遗传学等领域学生和研究人员的重要资源。立即开始你的学习之旅,探索自然界的隐秘真相!
One of the reasons why biotechnology has received so much attention during the past three decades is because of gene cloning. Although many useful products can be obtained from microbial culture, the list in the past has been limited to those compounds naturally synthesized by microorganisms. Many important pharmaceuticals, which are produced not by microbes but by higher organisms, could not be obtained in this way. This has been changed by the application of gene cloning to biotechnology.Gene cloning involves separating a specific gene or DNA segment from a larger chromosome, attaching it to a small molecule of carrier DNA, and then replicating this modified DNA thousands or millions of times through both an increase in host cell number and the creation of multiple copies of the cloned DNA in each cell.Particularly important to gene cloning is a set of enzymes made available through decades of research on nucleic acid metabolism. Two classes of enzymes lie at the heart of the classic approach to generating and propagating a recombinant DNA molecules i.e., Restriction nucleases and DNA ligase.Restriction endonucleases (also called restriction enzymes) recognize and cleave DNA at specific sequences (recognition sequences or restriction sites) to generate a set of smaller fragments. Some restriction endonucleases make staggered cuts on the two DNA strands, leaving two to four nucleotides of one strand unpaired at each resulting end.These unpaired strands are referred to as sticky ends. While, other restriction endonuclease cleave both strands of DNA at the opposing phosphodiester bonds, leaving no unpaired bases on the ends, often called blunt ends.The final step in construction of a recombinant DNA molecule is the joining together of the cloning vector and the gene to be cloned. This process is referred to as ligation, and the enzyme that catalyzes the reaction is called DNA ligase. A cloning vector is a small piece of DNA that can be stably maintained in an organism, and into which a foreign DNA fragment can be inserted for cloning purposes. A large number of cloning vectors are available. But, choosing the vector may depend upon a number of factors, such as the size of the insert, copy number and cloning method. Once a gene has been cloned there is almost no limit to the information that can be obtained about its structure and expression. The availability of cloned material has stimulated the development of analytical methods for studying genes, with new techniques being introduced all the time.Recombinant DNA technology has proven useful in attempting to cure these genetically inherited diseases. Gene therapy is the name originally given to methods that aim to cure an inherited disease by providing the patient with a correct copy of the defective gene. Gene therapy has now been extended to include attempts to cure any disease by introduction of a cloned gene into the patient. The clinical uses of gene therapy are not limited to treatment of inherited diseases. Most cancers result from activation of an oncogene that leads to tumor formation, or inactivation of a gene that normally suppresses formation of a tumor. In both cases a gene therapy could be envisaged to treat the cancer.This course is a valuable resource for students and researchers related to biology, forensic science, molecular biology, biotechnology, biochemistry and genetics. Start your learning journey now and explore the hidden truth about nature!