scholarly journals Gene Replacement in Gram-Negative Bacteria: the pMAKSAC Vectors

BioTechniques ◽  
2000 ◽  
Vol 28 (2) ◽  
pp. 198-204 ◽  
Author(s):  
Didier Favre ◽  
Jean-François Viret
Keyword(s):  
Gene Replacement ◽  
Gram Negative Bacteria ◽  
Gram Negative

scholarly journals Adaptation of the Yeast URA3 Selection System to Gram-Negative Bacteria and Generation of a ΔbetCDE Pseudomonas putida Strain

2005 ◽  
Vol 71 (2) ◽  
pp. 883-892 ◽  
Author(s):  
Teca Calcagno Galvão ◽  
Víctor de Lorenzo
Keyword(s):  
Pseudomonas Putida ◽  
General Procedure ◽  
Gene Replacement ◽  
Selection Marker ◽  
Homologous Gene ◽  
Gram Negative Bacteria ◽  
Selection System ◽  
Gram Negative ◽  
Acid Sensitivity ◽  
Delivery Vector

ABSTRACT A general procedure for efficient generation of gene knockouts in gram-negative bacteria by the adaptation of the Saccharomyces cerevisiae URA3 selection system is described. A Pseudomonas putida strain lacking the URA3 homolog pyrF (encoding orotidine-5′-phosphate decarboxylase) was constructed, allowing the use of a plasmid-borne copy of the gene as the target of selection. The delivery vector pTEC contains the pyrF gene and promoter, a conditional origin of replication (oriR6K), an origin of transfer (mobRK2), and an antibiotic selection marker flanked by multiple sites for cloning appropriate DNA segments. The versatility of pyrF as a selection system, allowing both positive and negative selection of the marker, and the robustness of the selection, where pyrF is associated with uracil prototrophy and fluoroorotic acid sensitivity, make this setup a powerful tool for efficient homologous gene replacement in gram-negative bacteria. The system has been instrumental for complete deletion of the P. putida choline-O-sulfate utilization operon betCDE, a mutant which could not be produced by any of the other genetic strategies available.

scholarly journals pET expression vector customized for efficient seamless cloning

BioTechniques ◽  
2020 ◽  
Vol 69 (5) ◽  
pp. 384-387
Author(s):  
Dragana Dobrijevic ◽  
Lily A Nematollahi ◽  
Helen C Hailes ◽  
John M Ward
Keyword(s):  
Expression Vector ◽  
Gene Replacement ◽  
Expression Vectors ◽  
Gram Negative Bacteria ◽  
Golden Gate ◽  
Gram Negative ◽  
Large Numbers ◽  
Parallel Cloning ◽  
Natural Enzyme

Here we present a modification of the widely used pET29 expression vector for use in rapid and straightforward parallel cloning via a gene replacement and Golden Gate strategy. The modification can be applied to other expression vectors for Gram-negative bacteria. We have used the modified vectors to clone large numbers of bacterial natural enzyme variants from genomic and metagenomic sources for applications in biocatalysis.

scholarly journals An improved version of suicide vector pKNG101 for gene replacement in Gram‐negative bacteria

1997 ◽  
Vol 23 (2) ◽  
pp. 410-411 ◽  
Author(s):  
Mahfuzur R. Sarker ◽  
Guy R. Cornelis
Keyword(s):  
Gene Replacement ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Suicide Vector

scholarly journals New Mobilizable Vectors Suitable for Gene Replacement in Gram-Negative Bacteria and Their Use in Mapping of the 3′ End of theXanthomonas campestris pv. campestrisgum Operon

1999 ◽  
Vol 65 (1) ◽  
pp. 278-282 ◽  
Author(s):  
Federico Katzen ◽  
Anke Becker ◽  
M. Verónica Ielmini ◽  
Cristian G. Oddo ◽  
Luis Ielpi
Keyword(s):  
Xanthomonas Campestris ◽  
Marker Gene ◽  
Gene Replacement ◽  
Gram Negative Bacteria ◽  
Double Crossover ◽  
Gram Negative ◽  
Marker Exchange

ABSTRACT We describe useful vectors to select double-crossover events directly in site-directed marker exchange mutagenesis in gram-negative bacteria. These vectors contain the gusA marker gene, providing colorimetric screens to identify bacteria harboring those sequences. The applicability of these vectors was shown by mapping the 3′ end of the Xanthomonas campestris gum operon, involved in biosynthesis of xanthan.

New Suicide Vector for Gene Replacement in Yersiniae and Other Gram-Negative Bacteria

Plasmid ◽  
1993 ◽  
Vol 29 (2) ◽  
pp. 160-163 ◽  
Author(s):  
Elzbieta Skrzypek ◽  
Pryce L. Haddix ◽  
Gregory V. Plano ◽  
Susan C. Straley
Keyword(s):  
Gene Replacement ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Suicide Vector

Versatile suicide vectors which allow direct selection for gene replacement in Gram-negative bacteria

Gene ◽  
1993 ◽  
Vol 127 (1) ◽  
pp. 15-21 ◽  
Author(s):  
Jürgen Quandt ◽  
Michael F. Hynes
Keyword(s):  
Gene Replacement ◽  
Direct Selection ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Selection For

Bacterial-Mucosal Cell Junctional Complexes

1974 ◽  
Vol 32 ◽  
pp. 144-145
Author(s):  
Roger C. Wagner
Keyword(s):  
Intestinal Wall ◽  
Rat Colon ◽  
Mucosal Cell ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Mucosal Cells ◽  
Mucosal Lining ◽  
Degenerative Alterations ◽  
Junctional Complexes ◽  
Intestinal Mucosal Cells

Bacteria exhibit the ability to adhere to the apical surfaces of intestinal mucosal cells. These attachments either precede invasion of the intestinal wall by the bacteria with accompanying inflammation and degeneration of the mucosa or represent permanent anchoring sites where the bacteria never totally penetrate the mucosal cells.Endemic gram negative bacteria were found attached to the surface of mucosal cells lining the walls of crypts in the rat colon. The bacteria did not intrude deeper than 0.5 urn into the mucosal cells and no degenerative alterations were detectable in the mucosal lining.

Rapid demonstration of septic or infectious arthritis due to Gram-negative bacteria

1992 ◽  
Vol 50 (1) ◽  
pp. 686-687
Author(s):  
Jacob S. Hanker ◽  
Paul R. Gross ◽  
Beverly L. Giammara
Keyword(s):  
Chronic Arthritis ◽  
Blood Cultures ◽  
Gram Negative Bacteria ◽  
Infectious Arthritis ◽  
Bacterial Arthritis ◽  
Gram Positive ◽  
Gram Negative ◽  
Gram Positive Bacteria

Blood cultures are positive in approximately only 50 per cent of the patients with nongonococcal bacterial infectious arthritis and about 20 per cent of those with gonococcal arthritis. But the concept that gram-negative bacteria could be involved even in chronic arthritis is well-supported. Gram stains are more definitive in staphylococcal arthritis caused by gram-positive bacteria than in bacterial arthritis due to gram-negative bacteria. In the latter situation where gram-negative bacilli are the problem, Gram stains are helpful for 50% of the patients; they are only helpful for 25% of the patients, however, where gram-negative gonococci are the problem. In arthritis due to gram-positive Staphylococci. Gramstained smears are positive for 75% of the patients.

Ultrastructure of periplasmic bodies in gram-negative bacteria

1990 ◽  
Vol 48 (3) ◽  
pp. 640-641
Author(s):  
Xie Nianming ◽  
Ding Shaoqing ◽  
Wang Luping ◽  
Yuan Zenglin ◽  
Zhan Guolai ◽  
...  
Keyword(s):  
Electron Microscope ◽  
Campylobacter Jejuni ◽  
Proteus Mirabilis ◽  
Shigella Flexneri ◽  
Morphological Description ◽  
Gram Negative Bacteria ◽  
Periplasmic Space ◽  
Clostridium Tetani ◽  
Gram Negative ◽  
Brucella Canis

Perhaps the data about periplasmic enzymes are obtained through biochemical methods but lack of morphological description. We have proved the existence of periplasmic bodies by electron microscope and described their ultrastructures. We hope this report may draw the attention of biochemists and mrophologists to collaborate on researches in periplasmic enzymes or periplasmic bodies with each other.One or more independent bodies may be seen in the periplasmic space between outer and inner membranes of Gram-negative bacteria, which we called periplasmic bodies. The periplasmic bodies have been found in seven species of bacteria at least, including the Pseudomonas aeroginosa. Shigella flexneri, Echerichia coli. Yersinia pestis, Campylobacter jejuni, Proteus mirabilis, Clostridium tetani. Vibrio cholerae and Brucella canis.

Dual stain kit for the microscopic gram classification of ocular infection bacteria

1993 ◽  
Vol 51 ◽  
pp. 248-249
Author(s):  
Jacob S. Hanker ◽  
Dale N. Holdren ◽  
Kenneth L. Cohen ◽  
Beverly L. Giammara
Keyword(s):  
Contact Lenses ◽  
Ocular Infection ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Gram Staining ◽  
Ocular Infections ◽  
Different Types ◽  
Culture Positive ◽  
Increased Susceptibility

Keratitis and conjunctivitis (infections of the cornea or conjunctiva) are ocular infections caused by various bacteria, fungi, viruses or parasites; bacteria, however, are usually prominent. Systemic conditions such as alcoholism, diabetes, debilitating disease, AIDS and immunosuppressive therapy can lead to increased susceptibility but trauma and contact lens use are very important factors. Gram-negative bacteria are most frequently cultured in these situations and Pseudomonas aeruginosa is most usually isolated from culture-positive ulcers of patients using contact lenses. Smears for staining can be obtained with a special swab or spatula and Gram staining frequently guides choice of a therapeutic rinse prior to the report of the culture results upon which specific antibiotic therapy is based. In some cases staining of the direct smear may be diagnostic in situations where the culture will not grow. In these cases different types of stains occasionally assist in guiding therapy.

Sign in / Sign up

Export Citation Format

Share Document