大肠杆菌菌株.docx

1、大肠杆菌菌株大肠杆菌菌株E. coli genotypesFrom OpenWetWareJump to: navigation, searchContentshide 1 Nomenclature & Abbreviations 2 Methylation Issues in E. coli 3 Commonly used strains o 3.1 AG1 o 3.2 AB1157 o 3.3 BL21(AI) o 3.4 BL21(DE3) o 3.5 BL21 (DE3) pLysS o 3.6 BNN93 o 3.7 BW26434, CGSC Strain # 7658 o 3.8

2、 C600 o 3.9 C600 hflA150 (Y1073, BNN102) o 3.10 D1210 o 3.11 DB3.1 o 3.12 DH1 o 3.13 DH5 o 3.14 DH10B (Invitrogen) o 3.15 DH12S (Invitrogen) o 3.16 DM1 (Invitrogen) o 3.17 ER2566 (NEB) o 3.18 ER2267 (NEB) o 3.19 HB101 o 3.20 IJ1126 o 3.21 IJ1127 o 3.22 JM83 o 3.23 JM101 o 3.24 JM103 o 3.25 JM105 o 3

3、.26 JM106 o 3.27 JM107 o 3.28 JM108 o 3.29 JM109 o 3.30 JM109(DE3) o 3.31 JM110 o 3.32 JM2.300 o 3.33 LE392 o 3.34 Mach1 o 3.35 MC1061 o 3.36 MC4100 o 3.37 MG1655 o 3.38 Rosetta(DE3)pLysS o 3.39 Rosetta-gami(DE3)pLysS o 3.40 RR1 o 3.41 STBL2 (Invitrogen) o 3.42 STBL4 o 3.43 SURE (Stratagene) o 3.44

4、SURE2 (Stratagene) o 3.45 TOP10 (Invitrogen) o 3.46 Top10F (Invitrogen) o 3.47 W3110 o 3.48 XL1-Blue (Stratagene) o 3.49 XL2-Blue (Stratagene) o 3.50 XL2-Blue MRF (Stratagene) o 3.51 XL1-Red (Stratagene) o 3.52 XL10-Gold (Stratagene) o 3.53 XL10-Gold KanR (Stratagene) 4 Other genotype information so

5、urces 5 References edit Nomenclature & Abbreviations A listed gene name means that gene carries a loss of function mutation, a 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, , e14, rac) are listed only

6、 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 w

7、ith 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 hasnt got the restriction system. mB/K+/- = The (B/K) defines the strain lineage. The +/- indicates whether the s

8、train has or hasnt 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

9、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 = muta

10、tion in alanine transporter; cannot use alanine as a carbon source dapD = mutation in succinyl diaminopimelate aminotransferase leads to succinate or (lysine + methionine) requirement ( ) = chromosomal deletion of genes between the listed genes (may include unlisted genes!) dam = adenine methylation

11、 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 4,85

12、1,348. 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 prepa

13、rations 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 = mutant more resistant to phage P1 infection galK/U =

14、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

15、 gyrase; conveys resistance to ccdB colicin gene product hflA150 = protease mutation stabilizing phage cII protein; high frequency of lysogenization by (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

16、 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) lacZM15 = partial deleti

17、on of the lacZ gene that allows complementation of the -galactosidase gene; required for blue/white selection on XGal plates. Deletes the amino portion of lacZ (aa 11-41). leuB = requires leucine lon = deletion of the lon protease malA = cannot metabolize maltose mcrA = Mutation eliminating restrict

18、ion 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

19、CmAG or GmAC mtlA = cannot metabilize mannitol (Mu) = Mu prophage present. Mu 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

20、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+ (80) = Cell carries the lambdoid prophage 80. A defective version of this phage carrying lacZM15 deletion is present in some strains. p

21、LysS = 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

22、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 re

23、peats 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)

24、 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 srl = cannot metabolize sorbitol supE = glnV supF = tyrT thi = requires thiamine thyA = r

25、equires 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 tr

26、xB = mutation in thioredoxin reductase; enhances disulphide bond formation in the cytoplasm tsx = outer membrane protein mutation conveying resistance to phage T6 and colicin K tyrT = suppression of amber (UAG) stop codons by insertion of tyrosine; needed for some phage infection such as gt11. ung1

27、= allows uracil to exist in plasmid DNA xyl-5 = blocked xylose metabolism edit Methylation Issues in E. coli Type I methylation systems: o E. coli K-12 restricts DNA which is not protected by adenine methylation at sites AA*CN6GTGC or GCA*CN6GTT, encoded by the hsdRMS genes(EcoKI). Deletions in thes

28、e genes removes either the restriction or methylation or both of these functions. o E. coli B derivative strains contain an hsdRMS system (EcoBI) restricting and protectiing the sequence TGA*N8TGCT or AGCA*N8TCA. The mcrA gene (carried on the e14 prophage) restricts DNA which is methylated in CmCWGG

29、 or mCG sequences (methylation by the dcm gene product). The mcrBC genes restrict RmC sequences. The mrr gene product restricts adenine methylated sequences at CAG or GAC sites. E. coli methylates the adenine in GATC (and the corresponding A on the opposite strand) with the dam gene product. M.EcoKI

30、I methylates the first A at the palindromic site ATGCAT (as well as the corresponding A on the opposite strand), see (Kossykh VG (2004) J. Bact 186: 2061-2067 PMID 15028690) Note that this article has been retracted; the retraction appears to center on textual plagarism, not experimental results. Th

31、e homology to AvaIII is real. I think I believe it. tk 20:28, 9 December 2005 (EST). Rich Roberts reports: We have tried ourselves to detect activity with this gene product and cannot detect any methyltransferase activity. In our case we used antibodies able to detect N6-methyladenine or N4 methylcytosine in DNA. The ones we have are very sensitive and should have been able to detect 5 methyl groups in the whole E. coli chromosome. Nothing was detected in an over expressing strain