scholarly journals A Mutagenesis Assay for Reporter Gene Screening Using Partially Degenerate Oligonucleotides of the Tandems NNT and NNC

2015 ◽  
Vol 2015 ◽  
pp. 1-7
Author(s):  
Huifen Xu ◽  
Cuilan Zhou ◽  
Andy K. Zhang ◽  
Wen Li ◽  
Jia Zhang ◽  
...  
Keyword(s):  
Pharmaceutical Industry ◽  
Candidate Gene ◽  
Resistance Gene ◽  
Reporter Gene ◽  
Specific Protein ◽  
High Tolerance ◽  
Nonsense Mutations ◽  
Gene Screening ◽  
Zeocin Resistance

Not all proteins are tolerable to mutations. Whether a specific protein can be a mutable target is of importance in the biotechnology and pharmaceutical industry. This study reported a novel mutagenesis assay using tandem NNT and NNC oligonucleotides to test the mutability of a candidate gene. These two tandem oligonucleotides avoid the risk of forming nonsense mutations and render flexibility of truncating or expanding the insertion size. As a reporter gene, ZeoR (zeocin resistance gene) was confirmed to have a high tolerance for mutagenesis by this new assay.

Fine mapping and candidate gene screening of the downy mildew resistance gene RPF1 in Spinach

2018 ◽  
Vol 131 (12) ◽  
pp. 2529-2541 ◽  
Author(s):  
Hongbing She ◽  
Wei Qian ◽  
Helong Zhang ◽  
Zhiyuan Liu ◽  
Xiaowu Wang ◽  
...  
Keyword(s):  
Candidate Gene ◽  
Downy Mildew ◽  
Resistance Gene ◽  
Fine Mapping ◽  
Downy Mildew Resistance ◽  
Mildew Resistance ◽  
Gene Screening

scholarly journals Fine Mapping and Candidate Gene Discovery of the Soybean Mosaic Virus Resistance Gene, Rsv4

2010 ◽  
Vol 3 (1) ◽  
Author(s):  
M. A. Saghai Maroof ◽  
Dominic M. Tucker ◽  
Jeffrey A. Skoneczka ◽  
Brian C. Bowman ◽  
Sucheta Tripathy ◽  
...  
Keyword(s):  
Candidate Gene ◽  
Mosaic Virus ◽  
Resistance Gene ◽  
Fine Mapping ◽  
Virus Resistance ◽  
Soybean Mosaic Virus ◽  
Gene Discovery ◽  
Virus Resistance Gene

scholarly journals Molecular dissection of resistance gene cluster and candidate gene identification of Pl17 and Pl19 in sunflower by whole-genome resequencing

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Guojia Ma ◽  
Qijian Song ◽  
William R. Underwood ◽  
Zhiwei Zhang ◽  
Jason D. Fiedler ◽  
...  
Keyword(s):  
Candidate Gene ◽  
Gene Cluster ◽  
Resistance Gene ◽  
Snp Markers ◽  
Reference Sequence ◽  
Whole Genome ◽  
Genome Resequencing ◽  
Density Mapping ◽  
Family Protein ◽  
Whole Genome Resequencing

Abstract Sunflower (Helianthus annuus L.) production is challenged by different biotic and abiotic stresses, among which downy mildew (DM) is a severe biotic stress that is detrimental to sunflower yield and quality in many sunflower-growing regions worldwide. Resistance against its infestation in sunflower is commonly regulated by single dominant genes. Pl17 and Pl19 are two broad-spectrum DM resistance genes that have been previously mapped to a gene cluster spanning a 3.2 Mb region at the upper end of sunflower chromosome 4. Using a whole-genome resequencing approach combined with a reference sequence-based chromosome walking strategy and high-density mapping populations, we narrowed down Pl17 to a 15-kb region flanked by SNP markers C4_5711524 and SPB0001. A prospective candidate gene HanXRQChr04g0095641 for Pl17 was identified, encoding a typical TNL resistance gene protein. Pl19 was delimited to a 35-kb region and was approximately 1 Mb away from Pl17, flanked by SNP markers C4_6676629 and C4_6711381. The only gene present within the delineated Pl19 locus in the reference genome, HanXRQChr04g0095951, was predicted to encode an RNA methyltransferase family protein. Six and eight SNP markers diagnostic for Pl17 and Pl19, respectively, were identified upon evaluation of 96 diverse sunflower lines, providing a very useful tool for marker-assisted selection in sunflower breeding programs.

Induction of Tolerance to FVIII in Hemophilic Mice by Delivery of Apoptotic Syngeneic Fibroblasts Expressing a FVIII Transgene.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 768-768
Author(s):  
Rui-Jun Su ◽  
Laura Stewart ◽  
Angela Epp ◽  
Steven W. Pipe ◽  
Neil C. Josephson
Keyword(s):  
T Cell ◽  
Factor Viii ◽  
Cell Line ◽  
Resistance Gene ◽  
Hemophilia A ◽  
Foamy Virus ◽  
Tolerance Induction ◽  
Apoptotic Cells ◽  
Control Vector ◽  
Zeocin Resistance

Abstract In response to treatment with factor concentrates approximately 30% of patients with severe hemophilia A develop high titer inhibitory antibodies to Factor VIII. Inhibitor formation represents a major obstacle in treating patients of hemophilia A. We are investigating ways to prevent and treat inhibitors, in the mouse model of hemophilia A, through tolerance induction by infusion of FVIII vector modified apoptotic syngeneic fibroblasts. We generated a fibroblast cell line from a tail snip of a 129Sv−FVIIIKO mouse. We then transduced these cells with a bi−cistronic foamy virus vector expressing both the full length FVIII coding sequence and the zeocin resistance gene. Production of FVIII by these modified cells was confirmed by ELISA on cell culture medium. A cell line transduced with a control vector expressing only the zeocin resistance gene was also generated. Both vector modified cell lines were maintained under selective pressure and induced to apoptose by UV irradiation just prior to infusion into 129Sv−FVIIIKO mice. Mice were treated with two infusions of UV−irradiated FVIII expressing or control fibroblasts at three different weekly cell doses: 1 x 107, 2 x 106, or 2 x 105. Ten days later they were inoculated with 4 weekly intravenous doses of 0.2 μg albumin free recombinant human FVIII (ADVATE). An additional set of control mice were not given any apoptotic cells prior to inoculation with ADVATE. Blood was collected to determine anti−FVIII inhibitor titers one week after the final ADVATE dose. The Bethesda titers in mice treated with apoptotic FVIII expressing fibroblasts, at all 3 cell doses, were 3–5 fold lower than mice that received no cells prior to intravenous FVIII challenge (p < 0.02 for all 3 cohorts, Welch’s t−test). A pair wise comparison with mice that received equivalent numbers of control apoptotic cells demonstrated significantly lower Bethesda titers in the mice given two weekly doses of 2 x106 FVIII expressing cells (436 ± 140BU versus 1230 ± 433BU, p = 0.04). In mice that received 1 x 107 and 2 x 105 apoptotic FVIII expressing cells there was a trend for the development of lower Bethesda titers when compared to mice that received an equivalent numbers of apoptotic cells modified by the control vector. Four months after treatment with ADVATE mice from the no cell group and both the 1 x 107 control and FVIII vector expressing groups were re−challenged with 4 additional weekly doses of ADVATE. T cell proliferation assays showed the highest stimulation index in mice that received no cells and the lowest index in mice that were treated with apoptotic FVIII expressing fibroblasts. Bethesda titer results from these re−challenged mice are currently pending. Our data show that anti−Factor VIII inhibitory antibody titers can be reduced by the prior infusion of apoptotic syngeneic cells stably transduced with a FVIII expressing foamy virus vector. Furthermore, the higher titers seen in mice treated with cells containing a control vector demonstrate that the effect is dependent on delivery of FVIII within the apoptotic cells. Moreover, the T cell stimulation data seen in the mice re−challenged 4 months after initial inoculation with FVIII indicate that the decrease in immune priming is due to the induction of durable tolerance. Future work will focus on elucidating the mechanisms of tolerance induction in this model and on combining apoptotic cell delivery with other immune modulators to optimize results.

Targeted Exome Sequencing of Deafness Genes After Failure of Auditory Phenotype-Driven Candidate Gene Screening

2015 ◽  
Vol 36 (6) ◽  
pp. 1096-1102 ◽  
Author(s):  
Bong Jik Kim ◽  
Ah Reum Kim ◽  
Gibeom Park ◽  
Woong Yang Park ◽  
Sun O Chang ◽  
...  
Keyword(s):  
Candidate Gene ◽  
Exome Sequencing ◽  
Gene Screening ◽  
Targeted Exome Sequencing

scholarly journals Candidate Gene Identification with SNP Marker-Based Fine Mapping of Anthracnose Resistance Gene Co-4 in Common Bean

PLoS ONE ◽  
2015 ◽  
Vol 10 (10) ◽  
pp. e0139450 ◽  
Author(s):  
Andrew J. Burt ◽  
H. Manilal William ◽  
Gregory Perry ◽  
Raja Khanal ◽  
K. Peter Pauls ◽  
...  
Keyword(s):  
Candidate Gene ◽  
Common Bean ◽  
Resistance Gene ◽  
Fine Mapping ◽  
Snp Marker ◽  
Gene Identification ◽  
Anthracnose Resistance ◽  
Candidate Gene Identification

scholarly journals Candidate-Gene Screening and Association Analysis at the Autism-Susceptibility Locus on Chromosome 16p: Evidence of Association at GRIN2A and ABAT

2005 ◽  
Vol 76 (6) ◽  
pp. 950-966 ◽  
Author(s):  
Gabrielle Barnby ◽  
Aaron Abbott ◽  
Nuala Sykes ◽  
Andrew Morris ◽  
Daniel E. Weeks ◽  
...  
Keyword(s):  
Candidate Gene ◽  
Association Analysis ◽  
Susceptibility Locus ◽  
Gene Screening ◽  
Autism Susceptibility
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