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1.鉴定。鉴定所培养的单菌落中是否含有重组质粒,或仅含有自身环化的质粒。如 前所述,前者电泳后会出现大小不等的两条DNA条带,一条为线性质粒载体的DNA,另 条为插入的目的基因;后者仅有一条载体的DNA条带。比如 DDPCR、 RACEPCR、细 胞库筛选、基因重组,等
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1.重组质粒的鉴定。当质粒的重组或其它载体重组后,通常会发生质粒的重组失 败,包括质粒的自身环化。因而要求进行筛选,把重组成功的质粒找出来。在目前常用的 质粒和其它载体中含有相应的抗生素抗性基因,一旦重组成功,质粒环化(包括自身环 化),抗生素抗性基因表达,被转化的大肠杆菌便具备抗相应抗生素的能力,可以含该抗生 素的培养基中生长传代,不然,重组失败,大肠杆菌便不能抵抗该抗生素而死亡
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1.组织学的主要研究方法。 2.组织学方法研究中医的现状
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◼ All DNA is recombinant DNA. ◼ Genetic exchange works constantly to blend and rearrange chromosomes, most obviously during meiosis, when homologous chromosomes pair prior to the first nuclear division. ◼ During this pairing, genetic exchange between the chromosomes occurs. This exchange, classically termed crossing over, is one of the results of homologous recombination
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TOPIC 1 Principles of Transcriptional Regulation [watch the animation] TOPIC 2 Regulation of Transcription Initiation: Examples from Bacteria (Lac operon, alternative s factors, NtrC,MerR, Gal rep, araBAD operon) TOPIC 3 Examples of Gene Regulation after Transcription Initiation (Trp operon) TOPIC 4 The Case of Phage λ: Layers of Regulation
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Similarity of regulation between eukaryotes and prokaryote 1.Principles are the same: • signals (信号), • activators and repressors (激活蛋白和阻 遏蛋白) • recruitment and allostery, cooperative binding (招募,异构和协同结合) 2. The gene expression steps subjected to regulation are similar, and the initiation of transcription is the most pervasively regulated step
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◼ What are the main challenges of translation and how do organisms overcome them? ◼ What is the organization of nucleotide sequence information in mRNA? ◼ What is the structure of tRNAs, and how do aminoacyl tRNA synthetases recognize and attach the correct amino acids to each tRNA? ◼ How does the ribosome orchestrate the translation process?
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Most of the eukaryotic genes are mosaic (嵌合体), consisting of intervening sequences separating the coding sequence ◼ Exons (外显子): the coding sequences ◼ Introns (内含子) : the intervening sequences ◼ RNA splicing: the process by which introns are removed from the pre￾mRNA. ◼ Alternative splicing (可变剪接): some pre-mRNAs can be spliced in more than one way , generating alternative mRNAs. 60% of the human genes are spliced in this manner
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Although DNA replication, repair, homologous recombination occur with high fidelity to ensure the genome identity between generations, there are genetic processes that rearrange DNA sequences and thus lead to a more dynamic genome structure Two classes of genetic recombination for DNA rearrangement: • Conservative site-specific recombination (CSSR): recombination between two defined sequence elements • Transpositional recombination (Transposition): recombination between specific sequences and nonspecific DNA sites
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(1)不离开自己的座位。(2)收起书和笔记,不可放 在桌面上。(3)只能看和写自己的试卷(4)老师宣布 停的时候,所有学生必须放下自己的笔,眼睛 向前看。 (5)老师宣布收试卷时,试卷必须快速由每排的最左 边传向右边,每排最右边的同学负责把试卷交给当堂助 教
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