Marker‐Free System Using Ribosomal Promoters Enhanced Xylose/Glucose Isomerase Production in Streptomyces rubiginosus

2019 ◽  
Vol 14 (11) ◽  
pp. 1900114 ◽  
Author(s):  
Xiaojie Wang ◽  
Zixin Deng ◽  
Tiangang Liu
Keyword(s):  
Glucose Isomerase ◽  
Free System ◽  
Marker Free ◽  
Streptomyces Rubiginosus

New trends towards a marker-free system for gait analysis

2000 ◽  
Author(s):  
Elodie F. Calais ◽  
Franck Marzani ◽  
Louis Legrand ◽  
Alain Jacquemard
Keyword(s):  
Gait Analysis ◽  
Free System ◽  
Marker Free

Analysis and tracking of human gait via a marker-free system

2000 ◽  
Author(s):  
Elodie F. Calais ◽  
Louis Legrand
Keyword(s):  
Human Gait ◽  
Free System ◽  
Marker Free

scholarly journals High-pressure small-angle neutron scattering studies of glucose isomerase conformation in solution

2009 ◽  
Vol 42 (3) ◽  
pp. 461-468 ◽  
Author(s):  
Ewa Banachowicz ◽  
Maciej Kozak ◽  
Adam Patkowski ◽  
Gerhard Meier ◽  
Joachim Kohlbrecher
Keyword(s):  
Neutron Scattering ◽  
Small Angle ◽  
Small Angle Neutron Scattering ◽  
Electrostatic Interactions ◽  
Xylose Isomerase ◽  
Glucose Isomerase ◽  
Secondary Standard ◽  
High Structural Stability ◽  
Streptomyces Rubiginosus ◽  
Sans Data

Small-angle neutron scattering (SANS) of solutions of glucose/xylose isomerase fromStreptomyces rubiginosuswas measured as a function of pressure. It is shown that the structure of the enzyme in solution as seen by SANS is practically the same as that in the crystal and does not change with pressure up to 150 MPa. This reflects the unusually high structural stability of this material, which makes it extremely interesting to use as a secondary standard for pressure-dependent SANS experiments. This lack of pressure dependence of the SANS data also indicates that any possible change in hydration of the protein induced by pressure is not visible in the SANS curves. An appropriate correction procedure must be used for the SANS data in order to account for the distortion of the intensity curve due to hard-sphere and electrostatic interactions. After this correction, the isomerase can be readily used as a secondary standard for SANS measurements.

scholarly journals A marker-free system for highly efficient construction of vaccinia virus vectors using CRISPR Cas9

2015 ◽  
Vol 2 ◽  
pp. 15035 ◽  
Author(s):  
Ming Yuan ◽  
Xuefei Gao ◽  
Louisa S Chard ◽  
Zarah Ali ◽  
Jahangir Ahmed ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Free System ◽  
Virus Vectors ◽  
Highly Efficient ◽  
Efficient Construction ◽  
Marker Free

scholarly journals Screening for recombinants of Crambe abyssinica after transformation by the pMF1 marker-free vector based on chemical selection and meristematic regeneration

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Weicong Qi ◽  
Iris E. M. Tinnenbroek-Capel ◽  
Elma M. J. Salentijn ◽  
Jan G. Schaart ◽  
Jihua Cheng ◽  
...  
Keyword(s):  
Gene Fusion ◽  
Negative Selection ◽  
Free System ◽  
Crambe Abyssinica ◽  
Stepwise Selection ◽  
Recombinant Cells ◽  
Extremely High Frequency ◽  
Marker Free ◽  
Dna Fragment ◽  
Chemical Selection

Abstract The T-DNA region of pMF1 vector of marker-free system developed by Wageningen UR, has Recombinase R-LBD gene fusion and nptII and codA gene fusion between two recombination sites. After transformation applying dexamethasone (DEX) can activate the recombinase to remove the T-DNA fragment between recombination sites. The recombinant ought to be selected on 5-fluorocytocine (5-FC) because of codA converting 5-FC into 5-fluorouracil the toxic. A PMF1 vector was transformed into hexaploid species Crambe abyssinica. Two independent transformants were chosen for DEX-induced recombination and later 5-FC selection. In contrast to earlier pMF1 experiments, the strategy of stepwise selection based on meristematic regeneration was engaged. After a long period of 5-FC selection, recombinants were obtained successfully, but most of the survivors were wildtype and non-recombinant. The results revealed when applying the PMF1 marker-free system on C. abyssinica, 1) Increasing in the DEX concentration did not correspondingly enhance the success of recombination; 2) both of the DEX-induced recombination and 5-FC negative selection were apparently insufficient which was leading to the extremely high frequency in chimerism occurring for recombinant and non-recombinant cells in tissues; 3) the strategy of stepwise selection based on meristem tissue regeneration was crucial for successfully isolating the recombinant germplasm from the chimera.

Contribution to a marker-free system for human motion analysis

2002 ◽  
Vol 11 (3) ◽  
pp. 381 ◽  
Author(s):  
Elodie Calais ◽  
Louis Legrand ◽  
Yvon Voisin ◽  
Alain Diou
Keyword(s):  
Motion Analysis ◽  
Human Motion ◽  
Human Motion Analysis ◽  
Free System ◽  
Marker Free

scholarly journals Regulated expression of site-specific DNA recombination for precision genetic engineering of apple

2005 ◽  
Author(s):  
John L. Norelli ◽  
Moshe Flaishman ◽  
Herb Aldwinckle ◽  
David Gidoni
Keyword(s):  
Genetic Engineering ◽  
Genomic Research ◽  
Fruit Crops ◽  
Selectable Markers ◽  
Free System ◽  
Inducible Promoters ◽  
Marker Excision ◽  
Marker Free ◽  
The Impact ◽  
Selection Of

Objectives: The original objectives of this project were to: 1) evaluate inducible promoters for the expression of recombinase in apple (USDA-ARS); 2) develop alternative selectable markers for use in apple to facilitate the positive selection of gene excision by recombinase (Cornell University); 3) compare the activity of three different recombinase systems (Cre/lox, FLP/FRT, and R/RS)in apple using a rapid transient assay (ARO); and 4) evaluate the use of recombinase systems in apple using the best promoters, selectable markers and recombinase systems identified in 1, 2 and 3 above (Collaboratively). Objective 2 was revised from the development alternative selectable markers, to the development of a marker-free selection system for apple. This change in approach was taken due to the inefficiency of the alternative markers initially evaluated in apple, phosphomannose-isomerase and 2-deoxyglucose-6-phosphate phosphatase, and the regulatory advantages of a marker-free system. Objective 3 was revised to focus primarily on the FLP/FRT recombinase system, due to the initial success obtained with this recombinase system. Based upon cooperation between researchers (see Achievements below), research to evaluate the use of the FLP recombinase system under light-inducible expression in apple was then conducted at the ARO (Objective 4). Background: Genomic research and genetic engineering have tremendous potential to enhance crop performance, improve food quality and increase farm profits. However, implementing the knowledge of genomics through genetically engineered fruit crops has many hurdles to be overcome before it can become a reality in the orchard. Among the most important hurdles are consumer concerns regarding the safety of transgenics and the impact this may have on marketing. The goal of this project was to develop plant transformation technologies to mitigate these concerns. Major achievements: Our results indicate activity of the FLP\FRTsite-specific recombination system for the first time in apple, and additionally, we show light- inducible activation of the recombinase in trees. Initial selection of apple transformation events is conducted under dark conditions, and tissue cultures are then moved to light conditions to promote marker excision and plant development. As trees are perennial and - cross-fertilization is not practical, the light-induced FLP-mediated recombination approach shown here provides an alternative to previously reported chemically induced recombinase approaches. In addition, a method was developed to transform apple without the use of herbicide or antibiotic resistance marker genes (marker free). Both light and chemically inducible promoters were developed to allow controlled gene expression in fruit crops. Implications: The research supported by this grant has demonstrated the feasibility of "marker excision" and "marker free" transformation technologies in apple. The use of these safer technologies for the genetic enhancement of apple varieties and rootstocks for various traits will serve to mitigate many of the consumer and environmental concerns facing the commercialization of these improved varieties.  

Glucose isomerase from Streptomyces rubiginosus – potential molecular weight standard for small-angle X-ray scattering

2005 ◽  
Vol 38 (3) ◽  
pp. 555-558 ◽  
Author(s):  
Maciej Kozak
Keyword(s):  
Molecular Weight ◽  
Synchrotron Radiation ◽  
Small Angle ◽  
Glucose Isomerase ◽  
Radius Of Gyration ◽  
X Ray ◽  
X Ray Scattering ◽  
Streptomyces Rubiginosus ◽  
Molecular Weight Standard ◽  
Ray Scattering

Stability of solutions of glucose isomerase from Streptomyces rubiginosus on long-term storage and on exposure to synchrotron radiation has been studied by the small-angle X-ray scattering (SAXS) method. The values of the radii of gyration and forward scattering do not change significantly on storage and on exposure to synchrotron radiation. The mean value of the radius of gyration characterizing glucose isomerase is R G = 3.27 ± 0.02 nm. For comparison, a SAXS study of monodispersive and aggregated bovine serum albumin (BSA) has been carried out. The results show that glucose isomerase could be a more stable molecular weight standard for SAXS than BSA.

scholarly journals Room-Temperature Structure of Xylitol-Bound Glucose Isomerase by Serial Crystallography: Xylitol Binding in the M1 Site Induces Release of Metal Bound in the M2 Site

2021 ◽  
Vol 22 (8) ◽  
pp. 3892
Author(s):  
Ki Hyun Nam
Keyword(s):  
Metal Binding ◽  
Room Temperature ◽  
Glucose Isomerase ◽  
Bound State ◽  
Industrial Applications ◽  
Octahedral Coordination ◽  
Temperature Structure ◽  
High Fructose ◽  
Distorted Octahedral ◽  
Streptomyces Rubiginosus

Glucose isomerase (GI) is an important enzyme that is widely used in industrial applications, such as in the production of high-fructose corn syrup or bioethanol. Studying inhibitor effects on GI is important to deciphering GI-specific molecular functions, as well as potential industrial applications. Analysis of the existing xylitol-bound GI structure revealed low metal occupancy at the M2 site; however, it remains unknown why this phenomenon occurs. This study reports the room-temperature structures of native and xylitol-bound GI from Streptomyces rubiginosus (SruGI) determined by serial millisecond crystallography. The M1 site of native SruGI exhibits distorted octahedral coordination; however, xylitol binding results in the M1 site exhibit geometrically stable octahedral coordination. This change results in the rearrangement of metal-binding residues for the M1 and M2 sites, the latter of which previously displayed distorted metal coordination, resulting in unstable coordination of Mg2+ at the M2 site and possibly explaining the inducement of low metal-binding affinity. These results enhance the understanding of the configuration of the xylitol-bound state of SruGI and provide insights into its future industrial application.

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