scholarly journals MoCloFlex: A Modular Yet Flexible Cloning System

2019 ◽  
Vol 7 ◽  
Author(s):  
Carlo A. Klein ◽  
Leonie Emde ◽  
Aaron Kuijpers ◽  
Patrick Sobetzko
Keyword(s):  
Cloning System

scholarly journals Identification of the Propionicin F Bacteriocin Immunity Gene (pcfI) and Development of a Food-Grade Cloning System for Propionibacterium freudenreichii

2007 ◽  
Vol 73 (23) ◽  
pp. 7542-7547 ◽  
Author(s):  
Dag Anders Brede ◽  
Sheba Lothe ◽  
Zhian Salehian ◽  
Therese Faye ◽  
Ingolf F. Nes
Keyword(s):  
Selection Marker ◽  
Multiple Cloning Site ◽  
Propionibacterium Freudenreichii ◽  
Expression Plasmid ◽  
Physiochemical Properties ◽  
Food Grade ◽  
Cloning System ◽  
Immunity Gene ◽  
Membrane Bound ◽  
High Level

ABSTRACT This report describes the first functional analysis of a bacteriocin immunity gene from Propionibacterium freudenreichii and its use as a selection marker for food-grade cloning. Cloning of the pcfI gene (previously orf5 [located as part of the pcfABC propionicin F operon]) rendered the sensitive host 1,000-fold more tolerant to the propionicin F bacteriocin. The physiochemical properties of the 127-residue large PcfI protein resemble those of membrane-bound immunity proteins from bacteriocin systems found in lactic acid bacteria. The high level of immunity conferred by pcfI allowed its use as a selection marker for plasmid transformation in P. freudenreichii. Electroporation of P. freudenreichii IFO12426 by use of the pcfI expression plasmid pSL102 and propionicin F selection (200 bacteriocin units/ml) yielded 107 transformants/μg DNA. The 2.7-kb P. freudenreichii food-grade cloning vector pSL104 consists of the pLME108 replicon, a multiple cloning site, and pcfI expressed from the constitutive PpampS promoter for selection. The pSL104 vector efficiently facilitated cloning of the propionicin T1 bacteriocin in P. freudenreichii. High-level propionicin T1 production (640 BU/ml) was obtained with the IFO12426 strain, and the food-grade propionicin T1 expression plasmid pSL106 was maintained by ∼91% of the cells over 25 generations in the absence of selection. To the best of our knowledge this is the first report of an efficient cloning system that facilitates the generation of food-grade recombinant P. freudenreichii strains.

scholarly journals AdEasy-based cloning system to generate tropism expanded replicating adenoviruses expressing transgenes late in the viral life cycle

Gene Therapy ◽  
2005 ◽  
Vol 12 (17) ◽  
pp. 1347-1352 ◽  
Author(s):  
M Lie-A-Ling ◽  
C T Bakker ◽  
J G Wesseling ◽  
P J Bosma
Keyword(s):  
Life Cycle ◽  
Viral Life Cycle ◽  
Cloning System

scholarly journals Construction of a self-cloning system in the unicellular green alga Pseudochoricystis ellipsoidea

2015 ◽  
Vol 8 (1) ◽  
Author(s):  
Yuki Kasai ◽  
Kohei Oshima ◽  
Fukiko Ikeda ◽  
Jun Abe ◽  
Yuya Yoshimitsu ◽  
...  
Keyword(s):  
Green Alga ◽  
Cloning System ◽  
Unicellular Green Alga

Establishment of an Expression Cloning System for CD4+ T Cell Epitopes

2001 ◽  
Vol 284 (5) ◽  
pp. 1140-1147 ◽  
Author(s):  
Shinji Fujii ◽  
Yasushi Uemura ◽  
Leo Kei Iwai ◽  
Masayuki Ando ◽  
Satoru Senju ◽  
...  
Keyword(s):  
T Cell ◽  
Cd4 T Cell ◽  
Expression Cloning ◽  
T Cell Epitopes ◽  
Cloning System

Development of a Self-Cloning System for Actinomadura verrucosospora and Identification of Polyketide Synthase Genes Essential for Production of the Angucyclic Antibiotic Pradimicin

1999 ◽  
Vol 65 (6) ◽  
pp. 2703-2709 ◽  
Author(s):  
Tohru Dairi ◽  
Yoshimitsu Hamano ◽  
Tamotsu Furumai ◽  
Toshikazu Oki
Keyword(s):  
Nucleotide Sequence ◽  
Polyketide Synthase ◽  
Selectable Marker ◽  
Plasmid Vector ◽  
Open Reading Frame ◽  
Antibiotic Biosynthesis ◽  
Nucleotide Sequence Analysis ◽  
Reading Frame ◽  
Cloning System ◽  
Open Reading Frame 1

ABSTRACT A self-cloning system for Actinomadura verrucosospora, a producer of the angucyclic antibiotic pradimicin A (PRM A), has been developed. The system is based on reproducible and reliable protoplasting and regeneration conditions for A. verrucosospora and a novel plasmid vector that consists of a replicon from a newly found Actinomadura plasmid and a selectable marker cloned from the Actinomadurastrain. The system has an efficiency of more than 105CFU/microgram of DNA. Using this system, we have cloned and identified the polyketide synthase (PKS) genes essential for PRM A biosynthesis from A. verrucosospora. Nucleotide sequence analysis of the 3.5-kb SalI-SphI fragment showed that ketosynthase subunits (open reading frame 1 [ORF1] and ORF2) of the essential PKS genes have strong similarities (59 to 89%) to those for angucyclic antibiotic biosynthesis.

A DNA cloning system for interspecies gene transfer in antibiotic-producing Streptomyces

Nature ◽  
1980 ◽  
Vol 284 (5756) ◽  
pp. 526-531 ◽  
Author(s):  
Mervyn Bibb ◽  
Janet L. Schottel ◽  
Stanley N. Cohen
Keyword(s):  
Gene Transfer ◽  
Dna Cloning ◽  
Cloning System
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