大肠杆菌基因型列表111

大肠杆菌基因型列表111
大肠杆菌基因型列表111

A listed gene name means that gene carries a loss of function mutation, a Δ preceding a gene name means the gene is deleted. If a gene is not listed, it is not known to be mutated. Prophages present in wt K-12 strains (F, λ, e14, rac) are listed only if ab sent. E. coli

B strains are naturally lon- and dcm-.

F- = Does not carry the F plasmid

F+ = Carries the F plasmid. The cell is able to mate with F- through conjugation.

F'[ ] = Carries an F plasmid that has host chromosomal genes on it from a previous recombination event. This cell can also mate with F- through conjugation. Chromosomal genes carried in the F plasmid are listed in brackets.

rB/K+/- = The (B/K) defines the strain lineage. The +/- indicates whether the strain has or hasn't got the restriction system.

mB/K+/- = The (B/K) defines the strain lineage. The +/- indicates whether the strain has or hasn't got the modification (methylation) system.

hsdS = Both restriction and methylation of certain sequences is deleted from the strain. If you transform DNA from such a strain into a wild type strain, it will be degraded.

hsdR = For efficient transformation of cloned unmethylated DNA from PCR amplifications

INV( ) = chromosomal inversion between locations indicated

ahpC = mutation to alkyl hydroperoxide reductase conferring disulfide reductase activity

ara-14 = cannot metabolize arabinose

araD = mutation in L-ribulose-phosphate 4-epimerase blocks arabinose metabolism

cycA = mutation in alanine transporter; cannot use alanine as a carbon source

dapD = mutation in succinyl diaminopimelate aminotransferase leads to succinate or (lysine + methionine) requirement

Δ( ) = chromosomal deletion of genes between the listed genes (may include unlisted genes!) dam = adenine methylation at GATC sequences abolished; high recombination efficiency; DNA repair turned on

dcm = cytosine methylation at second C of CCWGG sites abolished

deoR = regulatory gene that allows constitutive expression of deoxyribose synthesis genes; permits uptake of large plasmids. See Hanahan D, US Patent 4,851,348. ***This has been called into question, as the DH10B genome sequence revealed that it is deoR+. See Durfee08, PMID 18245285.

dnaJ = one of the chaparonins inactivated; stabilizes some mutant proteins

dut1 = dUTPase activity abolished, leading to increased dUTP concentrations, allowing uracil instead of thymine incorporation in DNA. Stable U incorporation requires ung gene mutation as well.

endA1 = For cleaner preparations of DNA and better results in downstream applications due to the elimination of non-specific digestion by Endonuclease I

(e14) = excisable prophage like element containing mcrA gene; present in K-12 but missing in many other strains

galE = mutations are associated with high competence, increased resistance to phage P1 infection, and 2-deoxygalactose resistance. galE mutations block the production of UDP-galactose, resulting in truncation of LPS glycans to the minimal, "inner core". The exceptional competence of DH10B/TOP10 is thought to be a result of a reduced interference from LPS in the binding and/or

uptake of transforming DNA. galE15 is a point mutation resulting in a Ser123 -> Phe conversion near the enzyme's active site. See van Die, et al. PMID 6373734, Hanahan, et al. PMID 1943786, and EcoSal ISBN 1555811647. --Dcekiert 16:56, 23 January 2008 (CST)

galk = mutants cannot metabolize galactose and are resistant to 2-deoxygalactose. galK16 is an IS2 insertion ~170bp downstream of the galK start codon. See EcoSal ISBN 1555811647. --Dcekiert 16:56, 23 January 2008 (CST)

galU = mutants cannot metabolize galactose

gor = mutation in glutathione reductase; enhances disulphide bond formation

glnV = suppression of amber (UAG) stop codons by insertion of glutamine; required for some phage growth

gyrA96 = mutation in DNA gyrase; conveys nalidixic acid resistance

gyrA462 = mutation in DNA gyrase; conveys resistance to ccdB colicin gene product

hflA150 = protease mutation stabilizing phage cII protein; high frequency of lysogenization by λ Δ(lac)X74 = Deletion of the entire lac operon as well as some flanking DNA.

lacIq or lacIQ = overproduction of the lac repressor protein; -35 site in promoter upstream of lacI is mutated from GCGCAA to GTGCAA

lacIQ1 = overproduction of the lac repressor protein; contains a 15 bp deletion to create optimal -35 site in promoter upstream of lacI

lacY = deficient in lactose transport; deletion of lactose permease (M protein)

lacZΔM15 = partial deletion of the lacZ gene that allows α complementation of the β-galactosidase gene; required for blue/white selection on XGal plates. Deletes the amino portion of lacZ (aa 11-41).

leuB = requires leucine

Δlon = deletion of the lon protease

malA = cannot metabolize maltose

mcrA = Mutation eliminating restriction of DNA methylated at the sequence CmCGG (possibly mCG). Carried on the e14 prophage (q.v.)

mcrB = Mutation eliminating restriction of DNA methylated at the sequence RmC

metB = requires methionine

metC = requires methionine

mrr = Mutation eliminating restriction of DNA methylated at the sequence CmAG or GmAC mtlA = cannot metabilize mannitol

(Mu) = Mu prophage present. Muδ means the phage is defective.

mutS - mutation inhibits DNA repair of mismatches in unmethylated newly synthesized strands nupG = same as deoR

ompT = mutation in outer membrane protein protease VII, reducing proteolysis of expressed proteins

(P1) = Cell carries a P1 prophage. Cells express the P1 restriction system.

(P2) = Cell carries a P2 prophage. Allows selection against Red+ Gam+ λ

(φ80) = Cell carries the lambdoid prophage φ80. A defective version of this phage car rying lacZM15 deletion (as well as wild-type lacI, lacYA, and flanking sequences) is present in some strains. The φ80 attachment site is just adjacent to tonB.

pLysS = contains pLysS plasmid carrying chloramphenicol resistance and phage T7 lysozyme, effective at attenuating activity of T7 RNA polymerase, for better inhibition of expression under

non-induced conditions. The sequence can be found here.

proA/B = requires proline

recA1 = For reduced occurrence of unwanted recombination in cloned DNA; cells UV sensitive, deficient in DNA repair

recA13 = as for recA1, but inserts less stable.

recBCD = Exonuclease V; mutation in RecB or RecC reduces general recombination by a factor of 100; impaired DNA repair; UV sensitive, easier propagation of inverted repeats

recJ Exonuclease involved in alternate recombination

relA = relaxed phenotype; permits RNA synthesis in absence of protein synthesis

rha = blocked rhamose metabolism

rnc = encodes RnaseIII (rnc-14 is a common null mutant)

rne = encodes RnaseE (rne-3071 is a common temperature sensitive mutant)

rpsL = mutation in ribosomal protein S12 conveying streptomycin resistance; also called strA sbcBC = ExoI activity abolished; usually present in recBC strains; recombination proficient, stable inverted repeats

sr1 = cannot metabolize sorbitol

supE = glnV

supF = tyrT

thi = requires thiamine

thyA = requires thymidine

Tn10 = transposon normally carrying Tetracycline resistance

Tn5 = transposon normally carrying Kanamycin resistance

tonA = Mutation in outer membrane protein conveying resistance to phage T1 and phage T5

traD = Mutation eliminating transfer factor; prevents transfer of F plasmid

trxB = mutation in thioredoxin reductase; enhances disulphide bond formation in the cytoplasm tsx = outer membrane protein mutation conveying resistance to phage T6 and colicin K

tryT = suppression of amber (UAG) stop codons by insertion of tyrosine; needed for some phage infection such as λgt11.

ung1 = allows uracil to exist in plasmid DNA

xyl-5 = blocked xylose metabolism

SmR = Streptomycin resistance

大肠杆菌基因型及遗传符号说明系列一DXY

大肠杆菌基因型及遗传符号说明系列一 点击次数:982 作者:佚名发表于:2009-09-27 00:00转载请注明来自丁香园 来源:丁香园 实验室的一般大肠杆菌拥有4288条基因,每条基因的长度约为950bp,基因间的平均间隔为118bp (基因Ⅷ)。E.coli基因组中还包含有许多插入序列,如λ-噬菌体片段和一些其他特殊组份的片段,这些插入的片段都是由基因的水平转移和基因重组而形成的,由此表明了基因组具有它的可塑造性。 利用大肠杆菌基因组的这种特性对其进行改造,使其中的某些基因发生突变或缺失,从而给大肠杆菌带来可以观察到的变化,这种能观察到的特征叫做大肠杆菌的表现型(Phenotype),把引起这种变化的基因构成叫做大肠杆菌的基因型(Genotype)。具有不同基因型的菌株表现出不同的特性。 分子克隆中常用的大肠杆菌及其遗传标记按Demerec等1966年提出的命名原则,采用的菌株所有的基因都假定处于野生型状态,除非在基因型上另外注明。 大肠杆菌基因型的表示方法(Demerec, et, al. 1966): 一、一般规则: 1、根据基因产物或其作用产物的英文名称的第一个字母缩写成3个小写斜体字母来表示。例如:D NA Adenine Methylase→dam。 2、不同的基因座,其中任何一个突变所产生的表型变化可能相同,其表示方法是在3个小写斜体字母后加上一个斜体大写字母来表示区别。例如:Recombination→recA、recB、recC。 3、突变位点应通过在突变基因符号后加不同数字表示。如supE44(sup基因座E的44位突变)。

如果不知道几个等位基因中哪一/几个发生了功能性突变,则用连字符“ -”代替大写字母,如trp-31。 4、细菌的基因型中应该包含关于其携带的质粒或附加体的的信息。这些符号包括菌株携带的质粒或附加体、质粒或附加体上的突变基因座和突变位点。其基因符号应与基因座的表示符号明显区别,符号的第一个字母大写、不斜体并位于括号内;质粒或附加体上的突变基因座和突变位点的基因符号的表示方法与染色体上突变基因座、突变位点的符号相同。 5、对于携带附加体的菌株的完整基因型描述应包括附加体的状态(游离或整合)。以F因子为例,F-:F因子缺失;F+:自主性F因子,不携带任何遗传可识别染色体片段;F':携带有遗传可识别细菌染色体片段的自主性F因子;Hfr:整合到染色体上的F因子(high frequency of recombination)。当这些质粒或噬菌体片段变异或缺失时,用()“或”/“等以区别。例如:/F' [traD3 6、proAB、lac I q、lacZ. M 15] 6、某个基因或某个领域缺失时,在其基因型前面加上“ ”表示。例如:lac-proAB基因缺失时它的基因型表示为(lac-proAB)。 7、由于某种基因的变异导致大肠杆菌可以明显观察到特征变化,有时也用其表现型代替基因型进行表示。例如:某些抗药性的获得或丧失,用如下方式表示:Streptomycin抗性→Str +或Str r,Ampicilli n敏感性→ Amp-。(第一个字母要大写,“+”或“r”表示有抗性,“-”表示无抗性或敏感)。 8、根据某些特异性蛋白的变异及其导致的结果变化进行表示。例如:TH2菌株上有一种基因型表示如下:hsdS20 (rB-、mB-),其中S20代表特异性识别蛋白发生变异,()中的rB-、mB-表示由于 S20的变异而导致B株来源的hsdR和hsdM的功能缺失。 9、蛋白质的名称与对应的基因或等位基因相同,但不用斜体,且首字母大写,如,UvrA、UvrB。 二、基因符号和意义(见表1)

大肠杆菌的基因型 Takara公司

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基因工程及其在大肠杆菌生产人干扰素中的应用

基因工程及其在大肠杆菌生产人干扰素中的应用 一、课程设计目的 了解工业生产中的新型育种技术并比较不同育种技术的优势; 学习理解基因工程育种技术及其操作原理; 研究基因工程育种技术在人干扰素生产中的创新。 二、课程设计题目描述与要求 本文介绍一种二十世纪七十年代发展起来的一种新型生物技术——基因工程,介绍其在育种中的应用。文中重点介绍了基因工程育种的一般步骤,以及近年来出现的运用基因工程进行定向育种的主要新技术:基因的定点突变,易错PCR,DAN重排及基因组重排。之后,应用基因工程育种技术重组大肠杆菌BL21(pBAI)生产人干扰素a2b, 通过优化补料分批培养时葡萄糖的流加策略,提高了hIFNa2b的表达量和表达速率。不同的葡萄糖流加方式有各自的优点,采用恒速流加葡萄糖的方式,hIFNa2b的表达量达到6 540 mg/L,高于目前已知文献中hIFNa2b的最高表达量5 200 mg/L。

三、课程设计报告内容 引言 基因工程是二十世纪七十年代发展起来的一种新型生物技术,其发展从根本上改变了生物技术的研究和开发应用模式。1972年美国的Berg和Jackson等人将猿猴病毒基因组SV 40DNA、λ噬菌体基因以及大肠杆菌半乳糖操纵子在体外重组获得成功。翌年,美国斯坦佛大学的Cohen和Boyer等人在体外构建出含有四环素和链霉素连个抗性基因的重组质粒分子,将之导入大肠杆菌后,该重组质粒得以稳定复制,并赋予受体细胞相应的抗生素抗性,由此宣告了基因工程的诞生。在二十世纪八十年代以来,随着大批大批成果的出现及应用,基因工程带来了一场新的革命。 利用这些技术,可以直接地、有针对性地在DNA分子水平上改造生物的遗传性状。通过转入外源基因,微生物和动、植物细胞可以产生出自身原来没有的蛋白质。同样,利用重组DNA技术,也可以使一些原来存在量极低但有重要工业或医学用途的小分子(抗生素)或蛋白质之外的大分子物质得以大量生产。特别是随着重组DNA技术的完善和发展,以基因水平为核心的现代分子定向育种技术越来越受到工业微生物育种学家的关注,并展示了良好的应用前景。 1、基因工程育种 基因工程育种是在基因水平上,运用人为方法将所需的某一供体生物的遗传物质提取出来,在离体条件下用适当的工具酶进行切割后,与载体连接,然后导入另一细胞,使外源遗传物质在其中进行正常复制和表达引,与前几种育种技术相比,基因工程育种技术是人们在分子生物学指导下的一种自觉的、能像工程一样可预先设计和控制的育种新技术,它可实现超远缘杂交,因而是最新最有前途的一种育种新技术。基因工程技术的全部过程一般包括目的基因DNA片段的取得、DNA片段与基因载体的体外连接、外源基因转入宿主细胞和目标基因的表达等主要环节。 1.1 基因工程育种的一般步骤是: (1)目的基因的获得:一般通过化学合成法、物理化学法(包括密度梯度离心法、单链酶法、分子杂交法)、鸟枪无性繁殖法、酶促合成法(逆转录法)、Norther

大肠杆菌基因工程菌常用类型

1、大肠杆菌DH5a菌株 DH5a是世界上最常用的基因工程菌株之一。由于DH5α是DNA酶缺陷型菌株,有利于基因克隆,保存质粒,但该菌株的蛋白酶没有缺陷,表达的蛋白容易被降解,因此通常不作为表达菌株。E.coli DH5a在使用pUC系列质粒载体转化时,可与载体编码的β-半乳糖苷酶氨基端实现α-互补。可用于蓝白斑筛选鉴别重组菌株。 基因型:F-,φ80dlacZΔM15,Δ(lacZYA-argF)U169,deoR,recA1,endA1,hsdR17(rk-,mk+),phoA,supE44,λ-,thi-1,gyrA96,relA1 2、大肠杆菌BL21(DE3) 菌株 该菌株用于高效表达克隆于含有噬菌体T7启动子的表达载体(如pET系列)的基因。T7噬菌体RNA聚合酶位于λ噬菌体DE3区,该区整合于BL21的染色体上。该菌适合表达非毒性蛋白。 基因型:F-,ompT,hsdS(rBB-mB-),gal,dcm(DE3) 3、大肠杆菌BL21(DE3) pLysS菌株 该菌株含有质粒pLysS,因此具有氯霉素抗性。PLysS含有表达T7溶菌酶的基因,能够降低目的基因的背景表达水平,但不干扰目的蛋白的表达。该菌适合表达毒性蛋白和非毒性蛋白。 基因型:F-,ompThsdS(rBB-mB-),gal,dcm(DE3,pLysS,Camr 4、大肠杆菌JM109菌株 该菌株在使用pUC系列质粒载体进行DNA转化或用M13 phage载体进行转染时,由于载体DNA产生的LacZa多肽和JM09编码的LacZΔM15进行α-互补,从而显示β-半乳糖苷酶活性,由此很容易鉴别重组体菌株。 基因型:recA1,endA1,gyrA96,thi-1,hsdR17,supE44,relA1,Δ(lac-proAB)/F’[traD36,proAB+,lacIq,lacZΔM15] 5、大肠杆菌TOP10菌株 该菌株适用于高效的DNA克隆和质粒扩增,能保证高拷贝质粒的稳定遗传。 基因型:F- ,mcrAΔ(mrr-hsd RMS-mcrBC),φ80 ,lacZΔM15,△lacⅩ74,recA1 ,araΔ139Δ(ara-leu)7697,galU,galK,rps,(Strr) endA1,nupG 6、大肠杆菌HB101菌株 该菌株遗传性能稳定,使用方便,适用于各种基因重组实验。 基因型:supE44,hsdS20(rB-mB-),recA13,ara-14,proA2,lacY1,galK2,rpsL20,xyl-5,mtl-1,leuB6,thi-1 7.XL10-Gold菌株:所制备的感受态细胞是目前转化效率最高的感受态细胞,缺失几乎所有已知的限制酶切系统;同时缺失核酸内切酶(endA),提高了质粒DNA的产量和质量;重组酶缺陷型(recA)减少插入片段的同源重组概率,保证了插入DNA的稳定性,提高感受态转化效率及大质粒转化能力的宿主菌基因型。

大肠杆菌的基因型

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大肠杆菌基因型列表111

A listed gene name means that gene carries a loss of function mutation, a Δ preceding a gene name means the gene is deleted. If a gene is not listed, it is not known to be mutated. Prophages present in wt K-12 strains (F, λ, e14, rac) are listed only if ab sent. E. coli B strains are naturally lon- and dcm-. F- = Does not carry the F plasmid F+ = Carries the F plasmid. The cell is able to mate with F- through conjugation. F'[ ] = Carries an F plasmid that has host chromosomal genes on it from a previous recombination event. This cell can also mate with F- through conjugation. Chromosomal genes carried in the F plasmid are listed in brackets. rB/K+/- = The (B/K) defines the strain lineage. The +/- indicates whether the strain has or hasn't got the restriction system. mB/K+/- = The (B/K) defines the strain lineage. The +/- indicates whether the strain has or hasn't got the modification (methylation) system. hsdS = Both restriction and methylation of certain sequences is deleted from the strain. If you transform DNA from such a strain into a wild type strain, it will be degraded. hsdR = For efficient transformation of cloned unmethylated DNA from PCR amplifications INV( ) = chromosomal inversion between locations indicated ahpC = mutation to alkyl hydroperoxide reductase conferring disulfide reductase activity ara-14 = cannot metabolize arabinose araD = mutation in L-ribulose-phosphate 4-epimerase blocks arabinose metabolism cycA = mutation in alanine transporter; cannot use alanine as a carbon source dapD = mutation in succinyl diaminopimelate aminotransferase leads to succinate or (lysine + methionine) requirement Δ( ) = chromosomal deletion of genes between the listed genes (may include unlisted genes!) dam = adenine methylation at GATC sequences abolished; high recombination efficiency; DNA repair turned on dcm = cytosine methylation at second C of CCWGG sites abolished deoR = regulatory gene that allows constitutive expression of deoxyribose synthesis genes; permits uptake of large plasmids. See Hanahan D, US Patent 4,851,348. ***This has been called into question, as the DH10B genome sequence revealed that it is deoR+. See Durfee08, PMID 18245285. dnaJ = one of the chaparonins inactivated; stabilizes some mutant proteins dut1 = dUTPase activity abolished, leading to increased dUTP concentrations, allowing uracil instead of thymine incorporation in DNA. Stable U incorporation requires ung gene mutation as well. endA1 = For cleaner preparations of DNA and better results in downstream applications due to the elimination of non-specific digestion by Endonuclease I (e14) = excisable prophage like element containing mcrA gene; present in K-12 but missing in many other strains galE = mutations are associated with high competence, increased resistance to phage P1 infection, and 2-deoxygalactose resistance. galE mutations block the production of UDP-galactose, resulting in truncation of LPS glycans to the minimal, "inner core". The exceptional competence of DH10B/TOP10 is thought to be a result of a reduced interference from LPS in the binding and/or

仔猪大肠杆菌基因工程疫苗

仔猪大肠杆菌病基因工程灭活疫苗 K88ac-ST1-LTB) 新生仔猪大肠杆菌病是由产肠毒素性大肠埃希氏菌(Enterotoxigenic E.coli ,ETEC)引起的一种高度接触性、急性、致死性腹泻,特征是排黄色或黄白色稀粪。临床上以1? 7 日龄新生仔猪下痢为主要特征,其中1—3 日龄最多见。个别耐过仔猪经较长时间才能正常生长,但病愈存活后其生长发育和生产性能指标受到严重影响。 大肠杆菌性腹泻在我国广泛流行,新生仔猪大肠杆菌性腹泻的发病率和死亡率在不同地区各有差异,发病率为5.69%?86.5%,死亡率为17.0%?73.0% 。本病是影响养猪业发展的主要疾病之一。 产肠毒素性大肠杆菌(ETEC)有两类致病因子:一类为黏附素(或称定居因子,起吸附固定作用,主要分为K88 K99 987P、F41等,其中以心为主。另一类为肠毒素(耐热性肠毒素ST和不耐热性肠毒素LT),是直接导致腹泻的因子。细菌通过黏附素固定在肠黏膜表面,大量繁殖后产生ST与LT两种肠毒素,引起剧烈腹泻,脱水、酸中毒、低血钾等。 辽宁益康生物股份有限公司科研人员经过大量的流行病学调查,病原菌的分离鉴定,致病因子的研究,从细菌致病机理出发,成功地研发出免疫谱宽,免疫效果好,使用安全的仔猪大肠杆菌基因工程疫苗,从根本上解决了新生仔猪大肠杆菌性腹泻免疫预防这一难题,现将有关情况报告如下。 一、疫苗免疫机理 妊娠母猪临产前进行大肠杆菌基因工程灭活疫苗免疫接种,产生三种抗体(抗ST1抗 体、抗LT抗体、抗K88抗体),仔猪出生后吮食初乳,获得母源抗体产生被动免疫,从而抵抗大肠杆菌在肠道定居增殖,有效中和肠毒素,获得保护力。 二、产品特点 1、具有良好的免疫原性。 针对致泻因子耐热肠毒素(ST)、不耐热肠毒素(LT)和主要黏附因子瓯,采用基因工程技术构建的GE-3菌株,可以有效表达K88ac-ST i-LT B融合蛋白,并辅以氢氧化铝胶佐剂,在保留了K88ac和LTB 良好免疫原性基础上,赋予了ST1免疫原性。 2、构建菌株优良稳定。通过对菌株的安全性、有效性、菌种限定代次及保存条件、疫苗保存条件及保 存期等 试验,证实了该菌株是一株良好的疫苗候选株。 3、该疫苗生产工艺科学、质量稳定。通过对菌株发酵培养的培养基、诱导剂及其诱导条件、通气量及 菌液灭活等进行了筛 选和优化,确定了工业化生产工艺,并通过安全性、效力检验等试验,批次间稳定。 4、本疫苗安全性可靠、无毒副作用。采用实验动物模型接种,田间与区域本动物接种等试验进行安 全性评价,表明本疫 苗无肠毒素活性、无致病性,妊娠母猪临床母猪全部存活,均无流产、死胎和胎儿畸形等现象,分娩正常,所产仔猪生长发育良好。 5、免疫效果确实。经仔猪免疫保护试验证实,灭活疫苗免疫初产怀孕母猪两次,新生仔猪吮食一天初 乳后,用大肠杆菌强毒C83902 (K88ac+、ST+和LT+)株攻击新生仔猪,均获得了较好的免疫保护:保护率为90~97.4%,比未免疫组高16.17% ,平均日增重提高40g 。 三、安全试验、效力试验及推广应用效果情况 1安全试验 (1)倍量接种安全试验

常用大肠杆菌及其基因型

Commonly used strains https://www.360docs.net/doc/273096627.html,/wiki/E._coli_genotypes 1.AG1 endA1 recA1 gyrA96 thi-1 relA1 glnV44 hsdR17(r K - m K +) 2.AB1157 thr-1, araC14, leuB6(Am), Δ(gpt-proA)62, lacY1, tsx-33, qsr'-0, glnV44(AS), galK2(Oc), LAM-, Rac-0, hisG4(Oc), rfbC1, mgl-51, rpoS396(Am), rpsL31(strR), kdgK51, xylA5, mtl-1, argE3(Oc), thi-1?Bachmann BJ: Derivation and genotypes of some mutant derivatives of Escherichia coli K-12. Escherichia coli and Salmonella typhimurium. Cellular and Molecular Biology (Edited by: F C Neidhardt J L Ingraham KB Low B Magasanik M Schaechter H E Umbarger). Washington, D.C., American Society for Microbiology 1987, 2:1190-1219. See CGSC#1157 3.BL21 E. coli B F- dcm ompT hsdS(r B - m B -) gal [malB+] K-12 (λS) ?The "malB region" was transduced in from the K-12 strain W3110 to make the strain Mal+λS. See Studier et al. (2009) J. Mol. Biol. 394(4), 653 for a discussion of the extent of the transfer. ?Stratagene E. coli Genotype Strains 4.BL21(AI) F– ompT gal dcm lon hsdS B (r B - m B -) araB::T7RNAP-tetA ?an E. coli B strain carrying the T7 RNA polymerase gene in the araB locus of the araBAD operon q. ?Transformed plasmids containing T7 promoter driven expression are repressed until L-arabinose induction of T7 RNA polymerase.

E.coli genotypes 大肠杆菌基因型手册

From OpenWetWare 1 Nomenclature & Abbreviations 2 Methylation Issues in E. coli 3 Commonly used strains 3.1 AG1 3.2 AB1157 3.3 BL21(AI) 3.4 BL21(DE3) 3.5 BL21 (DE3) pLysS 3.6 BNN93 3.7 BW26434, CGSC Strain # 7658 3.8 C600 3.9 C600 hflA150 (Y1073, BNN102) 3.10 CSH50 3.11 D1210 3.12 DB3.1 3.13 DH1 3.14 DH5α 3.15 DH10B (Invitrogen) 3.16 DH12S (Invitrogen) 3.17 DM1 (Invitrogen) 3.18 ER2566 (NEB) 3.19 ER2267 (NEB) 3.20 HB101 3.21 HMS174(DE3) 3.22 IJ1126 3.23 IJ1127 3.24 JM83 3.25 JM101 3.26 JM103 3.27 JM105 3.28 JM106 3.29 JM107 3.30 JM108 3.31 JM109 3.32 JM109(DE3) 3.33 JM110 3.34 JM2.300 3.35 LE392 3.36 Mach1 3.37 MC1061 3.38 MC4100 3.39 MG1655 3.40 OmniMAX2

3.41 Rosetta(DE3)pLysS 3.42 Rosetta-gami(DE3)pLysS 3.43 RR13.44 STBL2 (Invitrogen)3.45 STBL43.46 SURE (Stratagene)3.47 SURE2 (Stratagene)3.48 TOP10 (Invitrogen)3.49 Top10F' (Invitrogen)3.50 W31103.51 XL1-Blue (Stratagene)3.52 XL2-Blue (Stratagene)3.53 XL2-Blue MRF' (Stratagene)3.54 XL1-Red (Stratagene)3.55 XL10-Gold (Stratagene)3.56 XL10-Gold KanR (Stratagene)4 Other genotype information sources 5 References A listed gene name means that gene carries a loss of function mutation, a Δ preceding a gene name means the gene is deleted. If a gene is not listed, it is not known to be mutated. Prophages present in wt K-12 strains (F, λ, e14, rac) are listed only if absent. E. coli B strains are naturally lon- and dcm-. F - = Does not carry the F plasmid F + = Carries the F plasmid. The cell is able to mate with F - through conjugation. F'[ ] = Carries an F plasmid that has host chromosomal genes on it from a previous recombination event. This cell can also mate with F - through conjugation. Chromosomal genes carried in the F plasmid are listed in brackets. r B/K +/- = The (B/K) defines the strain lineage. The +/- indicates whether the strain has or hasn't got the restriction system. m B/K +/- = The (B/K) defines the strain lineage. The +/- indicates whether the strain has or hasn't got the modification (methylation) system. hsdS = Both restriction and methylation of certain sequences is deleted from the strain. If you transform DNA from such a strain into a wild type strain, it will be degraded. hsdR = For efficient transformation of cloned unmethylated DNA from PCR amplifications INV( ) = chromosomal inversion between locations indicated ahpC = mutation to alkyl hydroperoxide reductase conferring disulfide reductase activity ara-14 = cannot metabolize arabinose araD = mutation in L-ribulose-phosphate 4-epimerase blocks arabinose metabolism cycA = mutation in alanine transporter; cannot use alanine as a carbon source dapD = mutation in succinyl diaminopimelate aminotransferase leads to succinate or (lysine +methionine) requirement Δ( ) = chromosomal deletion of genes between the listed genes (may include unlisted genes!)dam = adenine methylation at GATC sequences abolished; high recombination efficiency; DNA repair turned on dcm = cytosine methylation at second C of CCWGG sites abolished 通常dam/dcm都是默认的,无需标注,只有dam -、dcm -才有必要标出来,那是被迫使用某些酶切位点时才用来扩增质粒的特殊菌株。

大肠杆菌BL21(DE3)和DH5α、DH10B

DH5a是一种常用于质粒克隆的菌株。E.coli DH5a在使用pUC系列质粒载体转化时,可与载体编码的β-半乳糖苷酶氨基端实现α-互补。可用于蓝白斑筛选鉴别重组菌株。 基因型:F-,φ80dlacZΔM15,Δ(lacZYA-argF)U169,deoR,recA1,endA1,hsdR17(rk-,mk+),phoA,supE44,λ-,thi-1,gyrA96,relA1 2:BL21(DE3) 菌株 该菌株用于高效表达克隆于含有噬菌体T7启动子的表达载体(如pET系列)的基因。T7噬菌体RNA聚合酶位于λ 噬菌体DE3区,该区整合于BL21的染色体上。该菌适合表达非毒性蛋白。 基因型:F-,ompT,hsdS(rBB-mB-),gal,dcm(DE3) 3:BL21(DE3) pLysS菌株 该菌株含有质粒pLysS,因此具有氯霉素抗性。PLysS含有表达T7溶菌酶的基因,能够降低目的基因的背景表达水平,但不干扰目的蛋白的表达。该菌适合表达毒性蛋白和非毒性蛋白。 基因型:F-,ompT hsdS(rBB-mB-),gal,dcm(DE3,pLysS ,Camr 4:JM109菌株 该菌株在使用pUC系列质粒载体进行DNA转化或用M13 phage载体进行转染时,由于载体DNA产生的LacZa多肽和JM09编码的LacZΔM15进行α-互补,从而显示β-半乳糖苷酶活性,由此很容易鉴别重组体菌株 基因型:recA1,endA1,gyrA96,thi-1,hsdR17,supE44,relA1,Δ(lac-proAB)/F’ 5:TOP10菌株 该菌株适用于高效的DNA克隆和质粒扩增,能保证高拷贝质粒的稳定遗传。 基因型:F- ,mcrAΔ(mrr-hsd RMS-mcrBC),φ80 ,lacZΔM15,△lacⅩ74,recA1 ,araΔ139Δ(ara-leu)7697,galU ,galK ,rps,(Strr) endA1,nupG 6:HB101菌株 该菌株遗传性能稳定,使用方便,适用于各种基因重组实验 基因型:supE44,hsdS20(rB-mB-),recA13,ara-14,proA2,lacY1,galK2,rpsL20,xyl-5,mtl-1,leuB6,thi-1

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