Journal: Toxins

Article Title: Modified Heat-Stable Toxins (hSTa) of Enterotoxigenic Escherichia coli Lose Toxicity but Display Antigenicity after Being Genetically Fused to Heat-Labile Toxoid LT(R192G)

doi: 10.3390/toxins3091146

Figure Lengend Snippet: Escherichia coli strains and plasmids used in this study.

Article Snippet: Vector pUC19 (Promega, Madison, WI, USA) was used to clone and express the wildtype and mutated STa genes, and expression vector pET28α (Invitrogen, Carlsbad, CA, USA) was used to express LT and STa toxoid fusions.

Techniques: Plasmid Preparation, Construct, Recombinant, Negative Control

Journal:

Article Title: Prevalence of, Antibody Response to, and Immunity Induced by Haemophilus ducreyi Hemolysin

doi:

Figure Lengend Snippet: Bacterial strains and plasmids used in this study

Article Snippet: E. coli BL21(DE3) Host for protein expression Novagen, Madison, Wis. H. aphrophilus ATCC 33389 Arnold Smith H. ducreyi 35000-TcA Strain 35000 with Tn 916 inserted in hhdB , nonhemolytic 50 H. ducreyi 35000-KmA Strain 35000 with Tn 1545-Δ3 inserted in hhdB , nonhemolytic 50 H. ducreyi 35000ΔAPC Strain 35000 with cat cassette in hhdA gene, nonhemolytic 54 H. haemoglobinophilus ATCC 19416 ATCC, Manassas, Va. H. haemolyticus ATCC 33390 Arnold Smith H. influenzae ATCC 33911 Arnold Smith H. influenzae Rd Marilyn Roberts H. parainfluenzae ATCC 33392 Arnold Smith H. paraphrophilus ATCC 29237 Arnold Smith H. segmis ATCC 33393 Arnold Smith T. equigenitalis ATCC 35865 Arnold Smith Plasmids pUC19 E. coli cloning vector, Ap r GibcoBRL pCR2.1 TA cloning vector for cloning PCR products, Amp r Kan r Invitrogen, San Diego, Calif. pET24+ Expression plasmid with C-terminal His tag sequence and inducible T7 promoter, no translation initiation signals, Kan r Novagen pET24a+ Expression plasmid with C-terminal His tag sequence and inducible T7 promoter, Kan r Novagen pBCKS Cm r plasmid derived from pUC19 with KS multiple cloning site Stratagene, La Jolla, Calif. pPT384-ETa hhdBA genes cloned into pET24a+ Sst I- Sal I This study pLSSK Shuttle vector with ori , sulA , and strA genes from pLS88 and multiple cloning sites and lacZ gene from pBluescript SK 54 pPT384 hhdBA gene region in pTZ18 in same orientation as lac promoter 50 pPT376BCKS 5.8-kb Bgl II fragment containing part of hhdB and all of hhdA cloned into pBCKS Sst I- Sal I 50 pETBABN hhdBA genes cloned into pET24+ Bam HI- Sal I This study pLSBAHis+ hhdBA genes cloned into pLSSK in same orientation as lac promoter.

Techniques: Plasmid Preparation, Cloning, Expressing, TA Cloning, Sequencing, Derivative Assay, Clone Assay

Journal: Nucleic Acids Research

Article Title: Extensive libraries of gene truncation variants generated by in vitro transposition

doi: 10.1093/nar/gkx030

Figure Lengend Snippet: Overview of method to generate random deletions in a target gene. (1) A transposition reaction was performed to obtain random insertions of the transposon into the target gene. (2) 5΄ and 3΄ fragment sub-libraries of the gene were amplified in two separate PCR reactions. In each reaction, one primer was complementary to the 5΄ or 3΄ constant regions of the target gene, and the other to a region on the transposon. (3) The digestion with BsaI created unique overhangs in each library complementary to unique overhangs in the linker DNA (pUC19*-fragment) to favor directional ligation. (4) Libraries were ligated to the linker DNA, which was free of MlyI sites. (5) The product of ligation was treated with MlyI to remove the transposon sequence. (6) Intramolecular blunt-end ligation joined the 5΄ and 3΄ terminal fragments of the gene. (7) This circular library was linearized by PCR with primers complementary to the termini of the parental gene. (8) The final library of deletion variants of the desired size range was isolated by gel electrophoresis.

Article Snippet: TOPO TA cloning Kit and pUC19 vector (2.68 kb) were purchased from Invitrogen.

Techniques: Amplification, Ligation, Sequencing, Isolation, Nucleic Acid Electrophoresis