scholarly journals Cloning and Expression of Serratia Marcescens Prodigiosin Gene in Ecoli XL1blue

2021 ◽  
Vol 20 (2) ◽  
pp. 102-111
Author(s):  
Kumarss Amini ◽  
◽  
Maryam Akbari ◽  
Keyword(s):  
Escherichia Coli ◽  
Serratia Marcescens ◽  
Pharmaceutical Products ◽  
Cloning And Expression ◽  
Dust Particles ◽  
Negative Bacterium ◽  
Phylogenetic Studies ◽  
Close Relatives ◽  
Tract Infections ◽  
Cloned Gene

Background and Objectives: Serratia is a gram-negative bacterium. The pigmentation property of Serratia Marcescens is used as a marker of dust particles in the environment and in the hospital. Today biopigments are also widely used in the manufacture and production of pharmaceutical products. Prodigiosin is a promising drug due to its reported properties of antifungal immunosuppressive and anti-proliferative activities. In the present study, cloning of pig gene- isolated from Serratia Marcescens in Ecoli XL1blue was performed. Subjects and Methods 60 Samples were taken from clinical sources of patients hospitalized with urinary tract infections in Saveh Hospitals. Serratia Marcescens were identified and isolated by different tests. The pig gene was cloned by T-A cloning using PTG-19 vector into the Escherichia coli XL1blue as host. Expression of cloned gene in recombinant colonies was evaluated by Real time PCR. The phylogenetic tree was plotted using clustalX and Mega5 software Results Screening of samples identified 12 isolates of Serratia Marcescens from then 4 isolates had pig gene. Expression of Pig gene in Escherichia coli XL1blue was confirmed by Real-Tima PCR. As a result of phylogenetic studies, some close relatives of serratia have been identified as candidates for further studies Conclusion Serratia Marcescens can be considered as a rich source of pigments with many applications and can be used as indigenous strains to produce Prodigiosin.

scholarly journals Cloning and expression of LTB in Escherichia coli and Bacillus subtilis

2013 ◽  
Vol 16 (1) ◽  
pp. 13-22 ◽  
Author(s):  
Trang Thi Phuong Phan ◽  
Anh Le Tuan Nguyen ◽  
Hoang Duc Nguyen
Keyword(s):  
Escherichia Coli ◽  
Bacillus Subtilis ◽  
Fusion Protein ◽  
Expression System ◽  
Pharmaceutical Products ◽  
Cloning And Expression ◽  
Gene Encoding ◽  
E Coli ◽  
B Subunit ◽  
The Common

LTB is the B subunit of heat labile toxins (LT) in Escherichia coli ETEC. This subunit is non-toxic but has a high immune response. Therefore, LTB is considered a suitable antigen for partial vaccine against the diarrhea caused by E. coli ETEC. The most important component of partial vaccine is antigen protein. Nowadays, with the advancement of recombinant protein technology, these antigens are mainly produced by the common bacterial expression system as E. coli. However, the recombinant proteins produced by E. coli are often miscellaneous with enterotoxins, which should be removed from pharmaceutical products. Thus, the production of antigen proteins in other expression system without endotoxins like Bacillus subtilis is in attention. We conducted the experiments of cloning and expressing LTB using a novel pHT plasmid that allow the protein to be expressed in both of E. coli and B. subtilis. We were successful to generate plasmid pHT326 and express the gene encoding for the fusion protein of LTB and LysSN-6xHis-TEV in B. subtilis and E. coli. The binding of fusion protein on the columns that have affinity with His-tag was confirmed. This result is about to be applied for the development of partial vaccine aganst the diarrhea as well as the development of some diagnostic kits for ETEC in food or medical waste and kits to detect antibodies against LTB in animals.

scholarly journals Molecular cloning and expression in Escherichia coli of a Trichoderma viride endo-beta-(1 6)-galactanase gene

2004 ◽  
Vol 377 (3) ◽  
pp. 749-755 ◽  
Author(s):  
Toshihisa KOTAKE ◽  
Satoshi KANEKO ◽  
Aya KUBOMOTO ◽  
Md. Ashraful HAQUE ◽  
Hideyuki KOBAYASHI ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Signal Sequence ◽  
Trichoderma Viride ◽  
Degree Of Polymerization ◽  
Glycoside Hydrolases ◽  
Cloning And Expression ◽  
Arabinogalactan Protein ◽  
Reading Frame ◽  
Cloned Gene

A gene encoding endo-β-(1→6)-galactanase from Trichoderma viride was cloned by reverse transcriptase–PCR and expressed in Escherichia coli. The gene contained an open reading frame consisting of 1437 bp (479 amino acids). The deduced amino acid sequence of the protein showed little similarity with other known glycoside hydrolases. A signal sequence (20 amino acids) was found at the N-terminal region of the protein and the molecular mass of the mature form was calculated to be 50.488 kDa. The gene product expressed in E. coli as a recombinant protein fused with thioredoxin and His6 tags had almost the same substrate specificity and mode of action as native enzyme purified from a commercial cellulase preparation of T. viride, i.e. recombinant enzyme endo-hydrolysed β-(1→6)-galacto-oligomers with a DP (degree of polymerization) higher than 3, and it could also hydrolyse α-l-arabinofuranosidase-treated arabinogalactan protein from radish. It produced β-(1→6)-galacto-oligomers ranging from DP 2 to at least 8 at the initial hydrolysis stage and galactose and β-(1→6)-galactobiose as the major products at the final reaction stage. These results indicate that the cloned gene encodes an endo-β-(1→6)-galactanase. As far as we know, this is the first time an endo-β-(1→6)-galactanase has been cloned.

Molecular Analysis of Uropathogenic E.coli Isolates from Urinary Tract Infections

2019 ◽  
Vol 19 (3) ◽  
pp. 322-326 ◽  
Author(s):  
Hassan Valadbeigi ◽  
Elham Esmaeeli ◽  
Sobhan Ghafourian ◽  
Abbas Maleki ◽  
Nourkhoda Sadeghifard
Keyword(s):  
Escherichia Coli ◽  
Polymerase Chain Reaction ◽  
Urinary Tract ◽  
Urinary Tract Infections ◽  
Multiplex Polymerase Chain Reaction ◽  
Virulence Genes ◽  
Uropathogenic Escherichia Coli ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Tract Infections

Introduction: The aim of the current study was to investigate the prevalence of virulence genes in uropathogenic Escherichia coli (UPEC) isolates in Ilam. Materials and Methods: For this purpose, a total of 80 UPEC isolates were collected for patients with UTIs during a 6 months period. The multiplex polymerase chain reaction (multiplex PCR) was used to detect the papEF, fimH, iucD, hlyA, fyuA, and ompT genes. Results: The prevalence of fimH, papEF, iucD, fyuA, hlyA, hlyA, and ompT genes were 87.5%, 47.5%, 60%, 67.5%, 27.5%, 47.5% and 71.2%, respectively. Among all of the isolates, 27 profiles were obtained. Conclusion: Our findings demonstrated that the most prevalence was found for fimH, and different distribution of virulence genes suggested different ability of pathogenicity.

scholarly journals Urinary Excretion and Bactericidal Activities of Gemifloxacin and Ofloxacin after a Single Oral Dose in Healthy Volunteers

2001 ◽  
Vol 45 (12) ◽  
pp. 3524-3530 ◽  
Author(s):  
Christoph K. Naber ◽  
Michaela Hammer ◽  
Martina Kinzig-Schippers ◽  
Christian Sauber ◽  
Fritz Sörgel ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Urinary Tract ◽  
Urinary Excretion ◽  
Enterococcus Faecalis ◽  
Urinary Tract Infections ◽  
Parent Drug ◽  
Oral Dosage ◽  
Tract Infections ◽  
Cumulative Excretion

ABSTRACT In a randomized crossover study, 16 volunteers (8 men, 8 women) received single oral doses of 320 mg of gemifloxacin and 400 mg of ofloxacin on two separate occasions in the fasting state to assess the urinary excretion and urinary bactericidal titers (UBTs) at intervals for up to 144 h. Ofloxacin showed higher concentrations in urine compared with those of gemifloxacin. The median (range) cumulative excretion of gemifloxacin was 29.7% (8.4 to 48.7%) of the parent drug administered, and median (range) cumulative excretion of ofloxacin was 84.3% (46.5 to 95.2%) of the parent drug administered. The UBTs, i.e., the highest twofold dilutions (with antibiotic-free urine as the diluent) of urine that were still bactericidal, were determined for a reference strain and nine uropathogens for which the MICs of gemifloxacin and ofloxacin were as follows:Escherichia coli ATCC 25922, 0.016 and 0.06 μg/ml, respectively; Klebsiella pneumoniae, 0.03 and 0.06 μg/ml, respectively; Proteus mirabilis, 0.125 and 0.125 μg/ml, respectively; Escherichia coli, 0.06 and 0.5 μg/ml, respectively; Pseudomonas aeruginosa, 1 and 4 μg/ml, respectively; Staphylococcus aureus, 0.008 and 0.25 μg/ml, respectively; Enterococcus faecalis, 0.06 and 2 μg/ml, respectively;Staphylococcus aureus, 0.25 and 4 μg/ml, respectively;Enterococcus faecalis, 0.5 and 32 μg/ml, respectively; and Staphylococcus aureus, 2 and 32 μg/ml, respectively. Generally, the UBTs for gram-positive uropathogens were higher for gemifloxacin than for ofloxacin and the UBTs for gram-negative uropathogens were higher for ofloxacin than for gemifloxacin. According to the UBTs, ofloxacin-resistant uropathogens (MICs, ≥4 mg/liter) should also be considered gemifloxacin resistant. Although clinical trials have shown that gemifloxacin is effective for the treatment of uncomplicated urinary tract infections, whether an oral dosage of 320 mg of gemifloxacin once daily is also adequate for the treatment of complicated urinary tract infections has yet to be confirmed.

scholarly journals Avian Pathogenic Escherichia coli (APEC): An Overview of Virulence and Pathogenesis Factors, Zoonotic Potential, and Control Strategies

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 467
Author(s):  
Dipak Kathayat ◽  
Dhanashree Lokesh ◽  
Sochina Ranjit ◽  
Gireesh Rajashekara
Keyword(s):  
Escherichia Coli ◽  
Current Status ◽  
Neonatal Meningitis ◽  
Clear Understanding ◽  
Zoonotic Potential ◽  
Secretion Systems ◽  
Avian Pathogenic Escherichia Coli ◽  
E Coli ◽  
Pathogenic Escherichia Coli ◽  
Tract Infections

Avian pathogenic Escherichia coli (APEC) causes colibacillosis in avian species, and recent reports have suggested APEC as a potential foodborne zoonotic pathogen. Herein, we discuss the virulence and pathogenesis factors of APEC, review the zoonotic potential, provide the current status of antibiotic resistance and progress in vaccine development, and summarize the alternative control measures being investigated. In addition to the known virulence factors, several other factors including quorum sensing system, secretion systems, two-component systems, transcriptional regulators, and genes associated with metabolism also contribute to APEC pathogenesis. The clear understanding of these factors will help in developing new effective treatments. The APEC isolates (particularly belonging to ST95 and ST131 or O1, O2, and O18) have genetic similarities and commonalities in virulence genes with human uropathogenic E. coli (UPEC) and neonatal meningitis E. coli (NMEC) and abilities to cause urinary tract infections and meningitis in humans. Therefore, the zoonotic potential of APEC cannot be undervalued. APEC resistance to almost all classes of antibiotics, including carbapenems, has been already reported. There is a need for an effective APEC vaccine that can provide protection against diverse APEC serotypes. Alternative therapies, especially the virulence inhibitors, can provide a novel solution with less likelihood of developing resistance.

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