基因工程学原理专业名词.docx

1、基因工程学原理专业名词第十九章 基因工程Chapter 19 Gene Engineering第一节 绪论Section 1 IntroductionDNA含有4种杂环碱基：嘌呤类有腺嘌呤（A）和鸟嘌呤（G），嘧啶类有胞嘧啶（C）和胸腺嘧啶（T）；而在RNA中尿嘧啶（U）替代了结构非常相似的胸腺嘧啶。In DNA, there are four heterocyclic bases: adenine (A) and guanine (G) are purines; cytosine (C) and thymine (T) are pyrimidines. In RNA, thymine is

2、replaced by the structurally very similar pyrimidine, uracil (U).核苷由碱基共价结合于戊糖分子的1位而构成。RNA中的戊糖为核糖，构成核糖核苷或简称核苷；而在DNA中戊糖为2脱氧核糖，形成2脱氧核糖核苷或简称脱氧核苷。碱基+糖分子=核苷。 A nucleoside consists of a base covalently bonded to the 1-position of a pentose sugar molecule. In RNA the sugar is ribose and the compounds are ri

3、bonucleosides, or just nucleosides,whereas in DNA it is 2-deoxyribose,and the nucleosides are named 2-deoxyribonucleosides,or just deoxynucleosides. Base+sugar=nucleoside. DNA分子通常以双螺旋形式存在。两条独立的反向平行的单链DNA分子以右手螺旋方式相互缠绕，糖-磷酸骨架在外，靠氢键和堆积力相互配对的碱基在内。腺嘌呤（A）与胸腺嘧啶（T）配对，鸟嘌呤（G）与胞嘧啶（C）配对。两条链是互补的，一条链决定了另一条链的序列。 D

4、NA most commonly occurs as a double helix. Two separate and antiparallel chains of DNA are wound around each other in a right-handed helical (coiled) path, with the sugar-phosphate backbones on the outside and the bases, paired by hydrogen bonding and stacked on each other, on the inside. Adenine pa

5、irs with thymine; guanine pairs with cytosine. The two chains are complementary; one specifies the sequence of the other. 可移动遗传因子mobile genetic element *基因gene *基因作用gene action 基因激活gene activation 基因活性gene activity *基因簇gene cluster *基因座gene locus *基因操作gene manipulation *基因标签gene tag 基因寻靶gene targeti

6、ng *遗传图genetic map *基因拷贝gene copy *基因克隆gene cloning *基因含量gene content *基因表达gene expression *基因表达系统gene expression system 基因融合gene fusion *基因定位gene localization *基因突变gene mutation 结瘤基因nodulin gene 分枝糊精branched dextrin *活性activity 生物碱alkaloid *抗体工程antibody engineering *生物信息学Bio-Informatics *生物信息biolog

7、ical information *数据库database *遗传标记genetic marker *基因组genome *基因组学genomics 跳跃基因jumping gene *开放阅读框open reading frame 第二节 DNA的提取与纯化Section 2 Extraction and Purification of DNA核酸中的芳香族碱基在260nm处具有最大光吸收。The aromatic bases of nucleic acids absorb light with a max of 260nm.核酸碱基的消光系数与它所处的环境有关。单一的核苷酸的吸收值比RNA

8、和单链DNA的吸收值最大，单链DNA又比双链DNA大。这种双链DNA相对于单链DNA吸收值减少的现象就称为减色效应。The extinction coefficient of nucleic acid bases depends on their environment. The absorbance of isolated nucleotides is greater than that of RNA and singe-stranded DNA, which is in turn greater than that of double-stranded DNA. Double-strand

9、ed DNA is hypochromic with respect to single-stranded DNA.核酸260nm处的光吸收可用于测定其浓度。当浓度为1mgml-1，光程为1cm时，双链DNA的A260=20，RNA和单链DNA的A260=25。对于RNA和单链DNA，A260的值取决于碱基的组成和二级结构。The absorbance at 260nm is used to determine the concentration of nucleic acids. At a concentration of 1mg ml-1 and 1 cm pathlength, doub

10、le-stranded DNA has A260=20.RNA and single-stranded DNA have A260=25. The values for RNA and single-stranded DNA depend on base composition and secondary structure.A260/A280比值可用于估计双链DNA样品的纯度。对于纯DNA，该比值为1.8；若比值大于1.8，则意味着RNA 污染；若比值小于1.8，则有蛋白质污染。The A260/ A280 ratio of a double-stranded DNA sample can

11、be used to assess its purity. For pure DNA, the value is 1.8. Values above 1.8 suggest RNA contamination and those below 1.8 suggest protein contamination.升高温度可使DNA和RNA变性。RNA随着温度升高而逐渐变性，双链DNA在某一确定温度“熔解”为单链DNA，这一温度表示为Tm，该值是DNA的（G+C）含量的函数。核酸的变性可以A260的变化来测定。Increased temperature can bring about the den

12、aturation of DNA and RNA. RNA denatures gradually on heating, but double-stranded DNA melts cooperatively to give single strands at a defined temperature, Tm which is a function of the G+C content of the DNA. Denaturation may be detected by the change in A260.*提取extraction *提取率extraction rate *纯化pur

13、ification 共价闭合环状DNAcovalently closed circular DNA *碱基置换base substitution *结合conjugation 循环测序法cycle sequencing 示差杂交differential hybridization DNA指纹图DNA finger-printing *表达图expression map 碱基插入base insertion *碱基堆积base stacking 假基因pseudogene 基体basal body 必需基因essential gene 畸形malformation 描图mapping *标记ma

14、rker 黑色素瘤melanoma 孟德尔遗传Mendelian inheritance 单体性monosomy 小鼠模型mouse model *突变mutation 功能性克隆表达functional cloning expression 富GC序列GC-rich sequence 多数tRNAmajor tRNA *主平板master plate *小卫星DNAmini-satellite DNA 核酸酶nuclease *蛋白-蛋白交互作用protein-protein interaction *基因缺失gene deletion 基因剔除gene knock-out 通用转录因子ge

15、neral transcription factor *基因组印记genomic imprinting 非组蛋白nonhistone protein 非组蛋白骨架nonhistone protein scaffold 随机扩增多态DNArandom amplified polymorphic DNA *密码子codon *RNA编辑RNA editing 亲和层析affinity chromatography *琼脂 agar *琼脂糖agarose 反向转运antiport 反义基因anti sense gene 反义核酸anti sense nucleic acid 肌动蛋白纤维actin

16、 filament *主动运输active transport 腺病毒adenovirus 粘着带adhesion belt 协助装配aided-assembly 选择性剪接alternate splicing 后期促进因子复合物anaphase-promoting complex 锚定连接anchoring junction 第三节 基因文库Section 3 Gene Library由基因组DNA所制成的基因文库称为基因组文库，而由互补DNA所制成的称为cDNA文库，两者统称为基因文库。cDNA文库不包括不能转录的核基因序列（重复序列等）。基因文库的好坏要看该文库是否覆盖起始材料的全部基因

17、或序列而未因克隆操作丧失其中的基因或某些序列。Gene libraries made from genomic DNA are called genomic libraries and those made from complementary DNA are known as cDNA libraries. The latter lack nontranscribed genomic sequences (repetitive sequences, etc.). Good gene libraries are representative of the starting material a

18、nd have not lost certain sequences due to cloning artifacts.基因文库必须包含一定数量的重组体，以便具有包含任何特定序列的可能性。如果知道基因组大小以及插入载体的片段的平均大小，则可以计算出所需重组体的数量。A gene library must contain a certain number of recombinants for there to be a high probability of it containing any particular sequence. This value can be calculated

19、if the genome size and the average size of the insert in the vector are known.为了构建文库，通常用蛋白酶降解及分相抽提制备基因组DNA，要用物理剪切或限制性酶解来进行随机分割，以形成适合于所用载体大小范围的片段。常用一组限制酶对DNA进行部分降解。For making libraries, genomic DNA, usually prepared by protease digestion and phase extraction, is fragmented randomly by physical sheari

20、ng or restriction enzyme digestion to give a size range appropriate for the chosen vector. Often combinations of restriction enzymes are used to partially digest the DNA.质粒、噬菌体、黏粒、BAC或酵母人工染色体等载体都可被用来构建基因组文库，选用那种载体取决于基因组的大小。这些载体所能插入的片段大小的上限分别依次为10kb、23kb、45kb、350kb和1000kb。用T4 DNA连接酶将基因组DNA片段与制备好的载体分子

21、相连接。Plasmids, phage, cosmid or yeast artificial chromosome vectors can be used to construct genomic libraries, the choice depending on the genome size. The upper size limit of these vectors is about 10,23,45 and 1000 kb respectively. The genomic DNA fragments are ligated to the prepared vector molec

22、ules using T4 DNA ligase.第一条链的合成是通过向引物，通常为寡聚（dT）的3端添加脱氧核糖核苷酸，用反转录酶来合成mRNA的cDNA拷贝。合成反应由示踪标记进行监测。用末端转移酶对cDNA第一链的3端进行加尾可以很容易合成出全长的第二链。反转录酶或Klenow酶都可与同聚物尾部例如，poly(dC)退火配对来延伸引物例如，寡聚（dG）来合成第二链cDNA。In first strand synthesis, reverse transcriptase is used to make a cDNA copy of the mRNA by extending a prime

23、r, usually oligo (dT), by the addition of deoxyribonucleotides to the 3-end. Synthesis can be detected by trace labeling. 3-Tailing of the first strand cDNA using terminal transferase makes full-length second strand synthesis easier. Reverse transcriptase or Klenow enzyme can extend a primer e.g. ol

24、igo (dG) annealed to a homopolymeric tail e.g. poly (dC) to synthesize second strand cDNA.*mRNA差异显示法mRNA differential display *基因组文库genomic library *基因文库gene library 核糖核酸ribonucleic acid 腺苷脱氨酶缺乏症adenosine deaminase deficiency 腺病毒adenovirus Alagille综合症Alagille syndrome *等位基因allele gene 动物模型animal model 抗体antibody 常染色体autosome 常染色体显性autosomal dominant Alu家族Alu family *自主性因子autonomous element *细菌人工染色体bacterial artificial chromosome *cDNA微矩阵分析cDNA micro-array analysis 顺反子cistron 共整合体cointegrate 组合库comb