A 3-D marker-free system for the analysis of movement disabilities - an application to the legs

2001 ◽  
Vol 5 (1) ◽  
pp. 18-26 ◽  
Author(s):  
F. Marzani ◽  
E. Calais ◽  
L. Legrand
Keyword(s):  
Free System ◽  
Marker Free

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 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.  

Combination of ribosome and phage display for fast selection of high affinity VHH antibody fragments

2019 ◽  
pp. 27-33
Author(s):  
YuE Kravchenko ◽  
SV Ivanov ◽  
DS Kravchenko ◽  
EI Frolova ◽  
SP Chumakov
Keyword(s):  
Phage Display ◽  
Selection Procedure ◽  
Free System ◽  
Phage Displays ◽  
High Affinity ◽  
Binding Domains ◽  
A Cell ◽  
Vhh Antibodies ◽  
Efficiency Of Selection ◽  
Selection Of

Selection of antibodies using phage display involves the preliminary cloning of the repertoire of sequences encoding antigen-binding domains into phagemid, which is considered the bottleneck of the method, limiting the resulting diversity of libraries and leading to the loss of poorly represented variants before the start of the selection procedure. Selection in cell-free conditions using a ribosomal display is devoid from this drawback, however is highly sensitive to PCR artifacts and the RNase contamination. The aim of the study was to test the efficiency of a combination of both methods, including pre-selection in a cell-free system to enrich the source library, followed by cloning and final selection using phage display. This approach may eliminate the shortcomings of each method and increase the efficiency of selection. For selection, alpaca VHH antibody sequences suitable for building an immune library were used due to the lack of VL domains. Analysis of immune libraries from the genes of the VH3, VHH3 and VH4 families showed that the VHH antibodies share in the VH3 and VH4 gene groups is insignificant, and selection from the combined library is less effective than from the VHH3 family of sequences. We found that the combination of ribosomal and phage displays leads to a higher enrichment of high-affinity fragments and avoids the loss of the original diversity during cloning. The combined method allowed us to obtain a greater number of different high-affinity sequences, and all the tested VHH fragments were able to specifically recognize the target, including the total protein extracts of cell cultures.

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