Journal:

Article Title: Cloning, Expression, and Characterization of the katG Gene, Encoding Catalase-Peroxidase, from the Polycyclic Aromatic Hydrocarbon-Degrading Bacterium Mycobacterium sp. Strain PYR-1

doi:

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

Article Snippet: Pyrene was obtained from Aldrich Chemical Company (Milwaukee, Wis.). table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Strain or plasmid Description Reference or source Strains Mycobacterium sp. strain PYR-1 Mineralizes high-molecular-weight PAHs such as fluoranthene, pyrene, and phenanthrene 8 Escherichia coli XL1-Blue MRF′ lac [F′ proAB lacI q ZΔM15 Tn 10 (Tet r )] Stratagene Escherichia coli XLOLR lac [F′ proAB lacI q ZΔM15 Tn 10 (Tet r )] Su − (nonsuppressing) λ r Stratagene Escherichia coli TOP10 One Shot competent cells, F′ mcrA Δ( mmr-hsdRMS-mcrBC ) Φ80 lacZ ΔM15 Δ( ara-leu ) 7697 Invitrogen Plasmid or vector ExAssist helper phage For in vivo excision of the pBK-CMV phagemid from the ZAP Express vector with E. coli XLOLR Stratagene Zap Express vector Lambda vector, prokaryotic and eukaryotic expression, in vivo excision of the pBK-CMV vector Stratagene pBK-CMV vector Neo r Kan r , ColE1 origin, lacZ , cytomegalovirus promoter Stratagene Myco-pBK-CMV phagemid pBK-CMV phagemid with 1.8-kb insert from Mycobacterium sp. strain PYR-1 This study pBAD/Thio-TOPO vector Amp r pMB1 origin, six-His-Thioredoxin open reading frame, arabinose induced Invitrogen katG -pBAD/Thio plasmid pBAD/ThioFusion with the katG gene of Mycobacterium strain PYR-1 This study Open in a separate window Bacterial strains and plasmids used in this study PAH induction of the Mycobacterium sp. cultures and two-dimensional (2D) gel electrophoretic analysis of recovered proteins.

Techniques: Plasmid Preparation, In Vivo, Expressing

Journal:

Article Title: Nocardia sp. Carboxylic Acid Reductase: Cloning, Expression, and Characterization of a New Aldehyde Oxidoreductase Family

doi: 10.1128/AEM.70.3.1874-1881.2004

Figure Lengend Snippet: Strains and plasmids used in this study

Article Snippet: Ampicillin (100 μg/ml) was incorporated into LB medium to select for recombinants, and isopropyl-β- d -thiogalactopyranoside (IPTG) (1 mM) and/or 5-bromo-4-chloro-3-indolyl-β- d -galactopyranoside (X-Gal) (80 μg/ml) were included for recombinant identification. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Strain or plasmid Relevant properties a Source or reference Strains Nocardia sp. strain NRRL 5646 Wild type 24 E. coli JM 109 RecA − , recombinant vector host strain Promega E. coli BL21 (DE3) Inducible T7 RNA polymerase, Amp r Stratagene E. coli BL21-CodonPlus (DE3)-RP Has argU and proL tRNA genes to provide tRNA for rare codons found in many GC-rich bacteria Stratagene Plasmids pGEM-T easy T/A PCR cloning vector, Amp r Promega pHAT10 Cloning vector for in-frame addition of HAT tag (MTMITPSLKDHLIHNVH KEEHAHAHNKIDDDDKVDGS) to the N terminus of CAR, pLac for expression, Amp r Clontech pHAT-305 pHAT-10 with car insert This study pHAT-DHFR Positive control expression vector with dihydrofolate reductase gene tagged with HAT at the N terminus Clontech Open in a separate window a Amp r , ampicillin resistance.

Techniques: Plasmid Preparation, Recombinant, PCR Cloning, Clone Assay, Expressing, Positive Control

Journal:

Article Title: Nocardia sp. Carboxylic Acid Reductase: Cloning, Expression, and Characterization of a New Aldehyde Oxidoreductase Family

doi: 10.1128/AEM.70.3.1874-1881.2004

Figure Lengend Snippet: SDS-PAGE and Western blot analysis of Nocardia CAR expression in E. coli carrying pHAT10-based vectors. Samples were taken from the lysates of E. coli cells carrying different vectors and were separated in duplicate by SDS-10% PAGE and either stained with 0.1% Coomassie blue R-250 (A) or subjected to Western blotting using a HAT-specific antibody (B). Lane 1, molecular mass markers for myosin (209 kDa), β-galactosidase (124 kDa), bovine serum albumin (80 kDa), ovalbumin (49.1 kDa), carbonic anhydrase (34.8 kDa), soybean trypsin inhibitor (21.5 kDa), lysozyme (20.6 kDa), and aprotinin (7.1 kDa). Lane 2, E. coli CodonPlus cells carrying pHAT-DHFR. Lane 3, E. coli BL21(DE3) cells carrying pHAT-305 (induced); lane 4, E. coli CodonPlus cells carrying pHAT-305 (induced). Lane 5, purified HAT-CAR. Lane 6, E. coli CodonPlus cells carrying pHAT10.

Article Snippet: Ampicillin (100 μg/ml) was incorporated into LB medium to select for recombinants, and isopropyl-β- d -thiogalactopyranoside (IPTG) (1 mM) and/or 5-bromo-4-chloro-3-indolyl-β- d -galactopyranoside (X-Gal) (80 μg/ml) were included for recombinant identification. table ft1 table-wrap mode="anchored" t5 TABLE 1. caption a7 Strain or plasmid Relevant properties a Source or reference Strains Nocardia sp. strain NRRL 5646 Wild type 24 E. coli JM 109 RecA − , recombinant vector host strain Promega E. coli BL21 (DE3) Inducible T7 RNA polymerase, Amp r Stratagene E. coli BL21-CodonPlus (DE3)-RP Has argU and proL tRNA genes to provide tRNA for rare codons found in many GC-rich bacteria Stratagene Plasmids pGEM-T easy T/A PCR cloning vector, Amp r Promega pHAT10 Cloning vector for in-frame addition of HAT tag (MTMITPSLKDHLIHNVH KEEHAHAHNKIDDDDKVDGS) to the N terminus of CAR, pLac for expression, Amp r Clontech pHAT-305 pHAT-10 with car insert This study pHAT-DHFR Positive control expression vector with dihydrofolate reductase gene tagged with HAT at the N terminus Clontech Open in a separate window a Amp r , ampicillin resistance.

Techniques: SDS Page, Western Blot, Expressing, Staining, Purification

Journal:

Article Title: Membrane Association and Kinase-Like Motifs of the RamC Protein of Streptomyces coelicolor

doi: 10.1128/JB.184.17.4920-4924.2002

Figure Lengend Snippet: Strains, plasmids, and primers used

Article Snippet: pGEX-5x-1 , N-terminal GST fusion expression vector (Amp r ) , , Amersham Pharmacia.

Techniques: Plasmid Preparation, Expressing