Identification of Novel Biofilm Genes in APEC by Tn5 Transposon Mutant Library

2021 ◽  
Author(s):  
Jiangang Hu ◽  
Yi Gu ◽  
Huiqi Lu ◽  
Fangheng Yu ◽  
Xiangpeng Niu ◽  
...  
Keyword(s):  
Mutant Library ◽  
Tn5 Transposon

High-Throughput Screening and Characterization of a High-Density Soybean Mutant Library Elucidate the Biosynthesis Pathway of Triterpenoid Saponins

2019 ◽  
Vol 60 (5) ◽  
pp. 1082-1097 ◽  
Author(s):  
Panneerselvam Krishnamurthy ◽  
Yukiko Fujisawa ◽  
Yuya Takahashi ◽  
Hanako Abe ◽  
Kentaro Yamane ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
High Density ◽  
Biosynthesis Pathway ◽  
Mutant Library ◽  
Triterpenoid Saponins

scholarly journals Tn5 transposon mutagenesis in Acidovorax citrulli for identification of genes required for pathogenicity on cucumber

2011 ◽  
Vol 61 (2) ◽  
pp. 364-374 ◽  
Author(s):  
J. Liu ◽  
S. Z. Luo ◽  
Q. Zhang ◽  
Q. H. Wang ◽  
J. F. Chen ◽  
...  
Keyword(s):  
Transposon Mutagenesis ◽  
Tn5 Transposon ◽  
Acidovorax Citrulli

scholarly journals Transposition of Tn5367 in Mycobacterium marinum, Using a Conditionally Recombinant Mycobacteriophage

2003 ◽  
Vol 185 (5) ◽  
pp. 1745-1748 ◽  
Author(s):  
Jan Rybniker ◽  
Martina Wolke ◽  
Christiane Haefs ◽  
Georg Plum
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Skin Infection ◽  
Model Organism ◽  
Transposon Mutagenesis ◽  
Mycobacterium Marinum ◽  
Close Relative ◽  
Mutant Library ◽  
Human Pathogen ◽  
Valid Model ◽  
Highly Efficient

ABSTRACT Mycobacterium marinum is a close relative of the obligate human pathogen Mycobacterium tuberculosis. As with M. tuberculosis, M. marinum causes intracellular infection of poikilothermic vertebrates and skin infection in humans. It is considered a valid model organism for the study of intracellular pathogenesis of mycobacteria. Low transformation efficiencies for this species have precluded approaches using mutant libraries in pathogenesis studies. We have adapted the conditionally replicating mycobacteriophage phAE94, originally developed as a transposon mutagenesis tool for M. tuberculosis, to meet the specific requirements of M. marinum. Conditions permissive for phage replication in M. tuberculosis facilitated highly efficient transposon delivery in M. marinum. Using this technique we succeeded in generating a representative mutant library of this species, and we conclude that TM4-derived mycobacteriophages are temperature-independent suicide vectors for M. marinum.

scholarly journals Construction of Signature-tagged Mutant Library in Mesorhizobium loti as a Powerful Tool for Functional Genomics

DNA Research ◽  
2008 ◽  
Vol 15 (5) ◽  
pp. 297-308 ◽  
Author(s):  
Y. Shimoda ◽  
H. Mitsui ◽  
H. Kamimatsuse ◽  
K. Minamisawa ◽  
E. Nishiyama ◽  
...  
Keyword(s):  
Functional Genomics ◽  
Mutant Library ◽  
Mesorhizobium Loti

Directed Evolution of The PobR Allosteric Transcription Factor To Generate A Biosensor For 4-Hydroxymandelic Acid

2021 ◽  
Author(s):  
YaoYao Liang ◽  
Juan Luo ◽  
Chenhao Yang ◽  
Shuning Guo ◽  
Bowen Zhang ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Transcription Factors ◽  
Directed Evolution ◽  
Missense Mutation ◽  
Hydroxybenzoic Acid ◽  
Ligand Specificity ◽  
Mutant Library

Abstract 4-Hydroxymandelic acid (HMA) is widely applied in pharmaceuticals, food and cosmetics. In this study, we aimed to develop an allosteric transcription factors (aTFs) based biosensor for HMA. PobR, an aTF for HMA analog 4-hydroxybenzoic acid, was used to alter its selectivity and create novel aTFs responsive to HMA by directed evolution. We established a PobR mutant library with a capacity of 550,000 mutants using error-prone PCR and Megawhop PCR. Through our screening, two mutants were obtained with responsiveness to HMA. Analysis of each missense mutation indicating residues 122-126 were involved in its PobR ligand specificity. These results showed the effectiveness of directed evolution in switching the ligand specificity of a biosensor and improving HMA production.

scholarly journals Genetic Determinants Involved in the Susceptibility of Pseudomonas aeruginosa to β-Lactam Antibiotics

2010 ◽  
Vol 54 (10) ◽  
pp. 4159-4167 ◽  
Author(s):  
Carolina Alvarez-Ortega ◽  
Irith Wiegand ◽  
Jorge Olivares ◽  
Robert E. W. Hancock ◽  
José Luis Martínez
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Detailed Analysis ◽  
Barrier Function ◽  
Mechanisms Of Action ◽  
Mutant Library ◽  
Genetic Determinants ◽  
Intrinsic Resistance ◽  
Resistance Phenotype ◽  
Mixed Phenotype ◽  
Alginate Biosynthesis

ABSTRACT The resistome of P. aeruginosa for three β-lactam antibiotics, namely, ceftazidime, imipenem, and meropenem, was deciphered by screening a comprehensive PA14 mutant library for mutants with increased or reduced susceptibility to these antimicrobials. Confirmation of the phenotypes of all selected mutants was performed by Etest. Of the total of 78 confirmed mutants, 41 demonstrated a reduced susceptibility phenotype and 37 a supersusceptibility (i.e., altered intrinsic resistance) phenotype, with 6 mutants demonstrating a mixed phenotype, depending on the antibiotic. Only three mutants demonstrated reduced (PA0908) or increased (glnK and ftsK) susceptibility to all three antibiotics. Overall, the mutant profiles of susceptibility suggested distinct mechanisms of action and resistance for the three antibiotics despite their similar structures. More detailed analysis indicated important roles for novel and known β-lactamase regulatory genes, for genes with likely involvement in barrier function, and for a range of regulators of alginate biosynthesis.

scholarly journals Mannuronan C-5 Epimerases Suited for Tailoring of Specific Alginate Structures Obtained by High-Throughput Screening of an Epimerase Mutant Library

2013 ◽  
Vol 14 (8) ◽  
pp. 2657-2666 ◽  
Author(s):  
Anne Tøndervik ◽  
Geir Klinkenberg ◽  
Finn L. Aachmann ◽  
Britt Iren Glærum Svanem ◽  
Helga Ertesvåg ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Mutant Library
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