Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors

Yanisch-Perron, Celeste R.; Vieira, Jeffrey; Messing, Joachim

doi:10.1016/0378-1119(85)90120-9

Cited by 15,038 publications

(5,554 citation statements)

References 24 publications

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“…The E. coli/Fransicella shuttle vector known as pCU18 was created by the combination of the pUC18 vector (Yanischperron et al, 1985) commonly used in E. coli and pC194 a broad host range plasmid from Staphylococcus aureus (Horinouchi and Weisblum, 1982). Each parent plasmid was digested with NdeI and SphI and the resulting fragments were purified using QIAquick Gel Extraction Kit (Catalog number 27106; Qiagen, Valencia CA).…”

Section: Plasmid Constructionmentioning

confidence: 99%

“…The pCUG vectors were created by combining the promoterless green fluorescent protein (GFP) gene from pFPV25 (Valdivia and Falkow, 1996), the broad host range origin of replication and chloramphenicol cassette from Staphylococcus aureus pC194 (Horinouchi and Weisblum, 1982) with the origin of replication from pUC18 (Yanischperron et al, 1985). The GFP gene was digested from pFPV25 by using restriction enzymes EcoRI and HindIII.…”

Section: Plasmid Constructionmentioning

confidence: 99%

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Development of novel plasmid vectors and a promoter trap system in Francisella tularensis compatible with the pFLN10 based plasmids

Rasko

Esteban

Sperandio³

2007

Plasmid

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Francisella tularensis is a category A bioterror pathogen which in some cases can cause a severe and fatal human infection. Very few virulence factors are known in this species due to the difficulty in working with it as well as the lack of tools for genetic manipulation. This work describes the construction of a shuttle vector that can replicate in Escherichia coli and F. tularensis as well as two distinct promoter trap constructs based on the shuttle vector backbone. Replication in F. tularensis is based on the promiscuous origin of replication from the Staphylococcus aureus plasmid pC194. We demonstrate the novel plasmids can coexist with established F. tularensis vectors based on the pFNL10 plasmid, the current workhorse of F. tularensis genetics. Our promoter trap can identify promoters that are activated during intracellular growth and survival. These new vectors provide additional tools for the genetic manipulation of F. tularensis.

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Section: Plasmid Constructionmentioning

confidence: 99%

Section: Plasmid Constructionmentioning

confidence: 99%

Development of novel plasmid vectors and a promoter trap system in Francisella tularensis compatible with the pFLN10 based plasmids

Rasko

Esteban

Sperandio³

2007

Plasmid

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“…The DNA fragment for lacP, lacZ ␣ and polylinker was excised from pUC19 (Yanisch-Perron et al 1985) as a 468-bp VspI (AseI)-HgaI fragment, and was treated with Klenow. The resulting blunt-ended fragment was inserted into pLG339 (Stoker et al 1982), between the EcoRI and BamHI sites (also K D Cromie et al treated with Klenow), replacing the 5 0 -terminal region of the tet gene.…”

Section: Construction Of Pkc200mentioning

confidence: 99%

“…The region encoding the Cterminus of ␤ and that downstream in the pKC301 and pKC302 constructs is schematically represented. The fusion of the first 137 amino acids of ␤ 0 with 104 amino acids of the LacZ␣ peptide (or 89 amino acids in a non-supE strain; Yanisch-Perron et al 1985) results in a pale blue colony morphology in the appropriate ␣-complementing host strain on medium containing X-gal and IPTG. The region amplified during PCR mutagenesis is shown, illustrating the introduction of the SacI site between rpoB and rpoC.…”

Section: Construction Of Pkc302mentioning

confidence: 99%

Trans‐dominant mutations in the 3′‐terminal region of the rpoB gene define highly conserved, essential residues in the β subunit of RNA polymerase: the GEME motif

Cromie¹,

Ahmad²,

Malik³

et al. 1999

Genes to Cells

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Background: The multimeric DNA-dependent RNA polymerases are widespread throughout nature. The RNA polymerase of Escherichia coli, which is the most well characterized, consists of a holoenzyme with subunit stoichiometry of ␣ 2 ␤␤ 0 . The ␤ subunit is conserved and has been implicated in all stages of transcription. The extreme C-terminus of the ␤ subunit, which includes two well-conserved sequence segments, contributes to the active centre and has been proposed to act in transcriptional termination. We describe a genetic system for further characterizing the role of the extreme Cterminus of the ␤ subunit of E. coli RNA polymerase. This involves random, PCR (Polymerase Chain Reaction)-mediated mutagenesis of the 3 0 region of rpoB encoding the C-terminal 116 amino acids of ␤, followed by the isolation and characterization of trans-dominant-negative mutations.

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“…coli DH5a cells: F, endA1, hsd R17 (rk" , ink+), supE44, thil, t, fecAl, gyrA96, relA1, o80dlacZ, AM15 (12) were used for propagation of plasmids and for expression of ~glucanase genes. The vectors comprised pBR322 (2) and pUC19 (30). The recombinant plasmid pEGl (6) carries an insert with the B. amyloliquefaciens [3- plasmid pUC19/34 (7).…”

Section: Strains Plasmids and Growth Mediamentioning

confidence: 99%

Hybrid bacillus endo-(1–3, 1–4)-β-glucanases: Construction of recombinant genes and molecular properties of the gene products

Borriss

Olsen

Thomsen

et al. 1989

Carlsberg Res. Commun.

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Hybrid 13-glucanase genes were constructed by the reciprocal exchange of the two halves of the isolated fl-glucanase genes from Bacillus amyloliquefaciens and B. macerans. The I]-glucanase hybrid enzyme 1 (HI) contains the 107 amino-terminal residues of mature B. amyloliquefaciens [i-glucanase and the 107 carboxyl-terminal amino acid residues of B. macerans ~glucanase. The reciprocal ~glucanase hybrid enzyme 2 (H2) consists of the 105 amino-terminal residues from the B. macerans enzyme and the carboxyl-terminal 107 amino acids from B. amyloliquefaciens. The biochemical properties of the two hybrid enzymes differ significantly from each other as well as from both parental ~glucanases.Hybrid ~glucanase H 1 exhibits increased thermostability in comparison to other ~glucanases, especially in an acidic environment. This hybrid enzyme has maximum activity between pH 5.6 and 6.6, whereas the oH-optimum for enzymatic activity of B. amyloliquefaciens ~glucanase was found to be at pH 6 to 7 and for B. macerans at pH 6.0 to 7.5. Hybrid enzyme 1 being more heat stable than both parental enzymes represents a case of intragenic heterosis.Hybrid 13-glucanase 2 (H2) was found to be more thermolabile than the naturally occurring ~ghicanases it was derived from and the pH-optimum for enzymatic activity was determined to be between pH 7 and pH 8. INTRODUCI'IONThe mixed linked ( 1-3, l-4)q3-glucans constitute the major part of the endosperm cell walls of cereals like oat and barley. They can cause severe problems in the brewing industry such as reduced yield of extract and lowered rates of wort separation or beer filtration. Remaining fl-glucans in the finished beer may lead to formation of hazes and gelatinous precipitates ( 11 ). Barley ( I-3,1-4)-~-glucanases (EC 3.2.1.73) are synthesized in the scutellum and the aleurone layer during the early stages of germiAbbreviations: aa = amino adds; amp = ampicillin; bgl = ~-glucanase gene; ~glucan = (1-3,1-4)-~-D-glncan; PAGE = polyacrylamide gel clectrophoresis; SDS ffi sodium dodecyl sulfate; tet ffi tetracycline; SP = signal peptide.Springer-Verlag

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Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mpl8 and pUC19 vectors

Cited by 15,038 publications

References 24 publications

Development of novel plasmid vectors and a promoter trap system in Francisella tularensis compatible with the pFLN10 based plasmids

Development of novel plasmid vectors and a promoter trap system in Francisella tularensis compatible with the pFLN10 based plasmids

Trans‐dominant mutations in the 3′‐terminal region of the rpoB gene define highly conserved, essential residues in the β subunit of RNA polymerase: the GEME motif

Hybrid bacillus endo-(1–3, 1–4)-β-glucanases: Construction of recombinant genes and molecular properties of the gene products

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