28 research outputs found
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A heterologous expression system for bovine lens transmembrane main intrinsic protein (MIP) in Nicotiana tabacum plants
We have developed a heterologous expression system for transmembrane lens main intrinsic protein (MIP) in Nicotiana tabacum plant tissue. A native bovine MIP26 amplicon was subcloned into an expression cassette under the control of a constitutive Cauliflower Mosaic Virus promoter, also containing a neomycin phosphotransferase operon. This cassette was transformed into Agrobacterium tumefaciens by triparental mating and used to infect plant tissue grown in culture. Recombinant plants were selected by their ability to grow and root on kanamycin-containing media. The presence of MIP in the plant tissues was confirmed by PCR, RT-PCR and immunohistochemistry. A number of benefits of this system for the study of MIP will be discussed, and also its application as a tool for the study of heterologously expressed proteins in general
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MASELTOV Deliverable D7.1.1: Incidental Learning Framework
This is the first steps towards creating an âIncidental learning frameworkâ, and it is a work in progress.
It is intended that the Incidental learning framework will facilitate the creation of technology rich learning opportunities for immigrants within cities. The framework will be a descriptive mechanism that permits analysis, a generative tool to support software system design, and it will facilitate the communication of learning design ideas both visually and textually. The framework focuses on incidental learning i.e. learning that is spontaneous and unplanned, in the knowledge domains of interest to the MASELTOV project including health care, culture, and language and information access. However, it provides links and triggers to structured and reflective learning to back up and deepen learning that happens incidentally.
This document describes the initial version of the Incidental Learning Framework, presents a few examples of its use, and describes some conclusions and recommendations for work towards the next version of the framework including
âą Using the framework to develop it,
âą Extending the literature review,
âą Gathering evidence about immigrants day-to-day lives,
âą Running design workshop(s) using the framework
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Fruit Development in Sweet Cherry
Fruits are an important source of vitamins, minerals and nutrients in the human diet. They also contain several compounds of nutraceutical importance that have significant antioxidant and anti-inflammatory roles, which can protect the consumer from diseases, such as cancer, and cardiovascular disease as well as having roles in reducing the build-up of LDL-cholesterol in blood plasma and generally reduce the risks of disease and age-related decline in health. Cherries contain high concentrations of bioactive compounds and minerals, including calcium, phosphorous, potassium and magnesium, and it is, therefore, unsurprising that cherry consumption has a positive impact on health. This review highlights the development of sweet cherry fruit, the health benefits of cherry consumption, and the options for increasing consumer acceptance and consumption
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Options for the generation of seedless cherry, the ultimate snacking product
Cherry fruit contain a large stone and seed, making processing of the fruit laborious and consumption by the consumer challenging, inconvenient to eat âon the moveâ and potentially dangerous for children. Availability of fruit lacking the stone and seed would be potentially transformative for the cherry industry since such fruit would be easier to process and would increase consumer demand because of the potential reduction in costs. This review will explore the background of seedless fruit, in the context of the ambition to produce the first seedless cherry, carry out an in-depth analysis of the current literature around parthenocarpy in fruit and discuss the available technology and potential for producing seedless cherry fruit as an âultimate snacking productâ for the 21st century
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Identification and distribution of novel badnaviral sequences integrated in the genome of cacao (Theobroma cacao)
Theobroma cacao is one of the most economically important tropical trees, being the source of chocolate. As part of an ongoing study to understand the diversity of the badnavirus complex, responsible for the cacao swollen shoot virus disease in West Africa, evidence was found recently of virus-like sequences in asymptomatic cacao plants. The present study exploited the wealth of genomic resources in this crop, and combined bioinformatic, molecular, and genetic approaches to report for the first time the presence of integrated badnaviral sequences in most of the cacao genetic groups. These sequences, which we propose to name eTcBV for endogenous Theobroma cacao bacilliform virus, varied in type with each predominating in a specific genetic group. A diagnostic multiplex PCR method was developed to identify the homozygous or hemizygous condition of one specific insert, which was inherited as a single Mendelian trait. These data suggest that these integration events occurred before or during the species diversification in Central and South America, and prior to its cultivation in other regions. Such evidence of integrated sequences is relevant to the management of cacao quarantine facilities and may also aid novel methods to reduce the impact of such viruses in this crop
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The novel RASSF6 and RASSF10 candidate tumour suppressor genes are frequently epigenetically inactivated in childhood leukaemias.
BACKGROUND: The Ras-association family (RASSF) of tumour suppressor genes (TSGs) contains 10 members that encode proteins containing Ras-association (RA) domains. Several members of the RASSF family are frequently epigenetically inactivated in cancer, however, their role in leukaemia has remained largely uninvestigated. Also, RASSF10 is a predicted gene yet to be experimentally verified. Here we cloned, characterised and demonstrated expression of RASSF10 in normal human bone marrow. We also determined the methylation status of CpG islands associated with RASSF1-10 in a series of childhood acute lymphocytic leukaemias (ALL) and normal blood and bone marrow samples. RESULTS: COBRA and bisulphite sequencing revealed RASSF6 and RASSF10 were the only RASSF members with a high frequency of leukaemia-specific methylation. RASSF6 was methylated in 94% (48/51) B-ALL and 41% (12/29) T-ALL, whilst RASSF10 was methylated in 16% (8/51) B-ALL and 88% (23/26) T-ALL. RASSF6 and RASSF10 expression inversely correlated with methylation which was restored by treatment with 5-aza-2'deoxycytidine (5azaDC). CONCLUSION: This study shows the hypermethylation profile of RASSF genes in leukaemias is distinct from that of solid tumours and represents the first report of inactivation of RASSF6 or RASSF10 in cancer. These data show epigenetic inactivation of the candidate TSGs RASSF6 and RASSF10 is an extremely frequent event in the pathogenesis of childhood leukaemia. This study also warrants further investigation of the newly identified RASSF member RASSF10 and its potential role in leukaemia.RIGHTS : This article is licensed under the BioMed Central licence at http://www.biomedcentral.com/about/license which is similar to the 'Creative Commons Attribution Licence'. In brief you may : copy, distribute, and display the work; make derivative works; or make commercial use of the work - under the following conditions: the original author must be given credit; for any reuse or distribution, it must be made clear to others what the license terms of this work are
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Identification of Cacao Mild Mosaic Virus (CaMMV) and Cacao Yellow Vein-Banding Virus (CYVBV) in Cocoa (Theobroma cacao) Germplasm
Cocoa, Theobroma cacao, is an important tropical perennial crop grown widely in the humid
tropics. The exchange of cocoa germplasm between germplasm collections and breeding centres is
vital for varietal development. Intermediate quarantine facilities, such as the International Cocoa
Quarantine Centre, Reading UK (ICQC-R) play a vital role in ensuring the transfer of germplasm
whilst minimising the risk of spreading pests and diseases. Current screening procedures combine
visual inspection and molecular techniques, which are effective in detecting Cocoa swollen shoot
virus (CSSV), a badnavirus, which causes severe losses but are restricted to West Africa. However,
the detection of latent or mild virus infections that produce no visual symptoms has been a challenge.
Recently two badnavirus species of cocoa producing mild symptoms, cacao mild mosaic virus
(CaMMV) and cacao yellow vein-banding virus (CYVBV), have been sequenced. Here, we report
new assays for the detection of these two species, for the first time in non-symptomatic accessions.
Evolutionary and bioinformatic analyses of the viruses suggest their most recent source was from
Trinidad, though there is historic evidence that these viruses may have their origin in South America
and then become widespread globally over the last century. We also report a novel colorimetric
Loop-mediated isothermal amplification (LAMP) assay for the detection of CYVBV. This simple and
accurate method could be employed in field virus testing
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Production of haploids and doubled haploids in oil palm
Background: Oil palm is the worldâs most productive oil-food crop despite yielding well below its theoretical
maximum. This maximum could be approached with the introduction of elite F1 varieties. The development of
such elite lines has thus far been prevented by difficulties in generating homozygous parental types for F1
generation.
Results: Here we present the first high-throughput screen to identify spontaneously-formed haploid (H) and
doubled haploid (DH) palms. We secured over 1,000 Hs and one DH from genetically diverse material and derived
further DH/mixoploid palms from Hs using colchicine. We demonstrated viability of pollen from H plants and
expect to generate 100% homogeneous F1 seed from intercrosses between DH/mixoploids once they develop
female inflorescences.
Conclusions: This study has generated genetically diverse H/DH palms from which parental clones can be selected
in sufficient numbers to enable the commercial-scale breeding of F1 varieties. The anticipated step increase in
productivity may help to relieve pressure to extend palm cultivation, and limit further expansion into biodiverse
rainforest
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Genomic investigation of the strawberry pathogen Phytophthora fragariae indicates pathogenicity is associated with transcriptional variation in three key races
The oomycete Phytophthora fragariae is a highly destructive pathogen of cultivated strawberry (Fragaria Ă ananassa), causing the root rotting disease, âred coreâ. The host-pathogen interaction has a well described gene-for-gene resistance relationship, but to date neither candidate avirulence nor resistance genes have been identified. We sequenced a set of American, Canadian, and United Kingdom isolates of known race type, along with three representatives of the closely related pathogen of the raspberry (Rubus idaeus), P. rubi, and found a clear population structure, with a high degree of nucleotide divergence seen between some race types and abundant private variation associated with race types 4 and 5. In contrast, between isolates defined as United Kingdom races 1, 2, and 3 (UK1-2-3) there was no evidence of gene loss or gain; or the presence of insertions/deletions (INDELs) or Single Nucleotide Polymorphisms (SNPs) within or in proximity to putative pathogenicity genes could be found associated with race variation. Transcriptomic analysis of representative UK1-2-3 isolates revealed abundant expression variation in key effector family genes associated with pathogen race; however, further long read sequencing did not reveal any long range polymorphisms to be associated with avirulence to race UK2 or UK3 resistance, suggesting either control in trans or other stable forms of epigenetic modification modulating gene expression. This work reveals the combined power of population resequencing to uncover race structure in pathosystems and in planta transcriptomic analysis to identify candidate avirulence genes. This work has implications for the identification of putative avirulence genes in the absence of associated expression data and points toward the need for detailed molecular characterisation of mechanisms of effector regulation and silencing in oomycete plant pathogens
The novel <i>RASSF6</i> and <i>RASSF10</i> candidate tumour suppressor genes are frequently epigenetically inactivated in childhood leukaemias
Background: The Ras-assocation family (RASSF) of tumour suppressor genes (TSGs) contains 10 members that encode proteins containing Ras-assocation (RA) domains. Several members of the RASSF family are frequently epigenetically inactivated in cancer, however, their role in leukaemia has remained largely uninvestigated. Also, RASSF10 is a predicted gene yet to be experimentally verified. Here we cloned, characterised and demonstrated expression of RASSF10 in normal human bone marrow. We also determined the methylation status of CpG islands associated with RASSF1-10 in a series of childhood acute lymphocytic leukaemias (ALL) and normal blood and bone marrow samples. Results: COBRA and bisulphite sequencing revealed RASSF6 and RASSF10 were the only RASSF members with a high frequency of leukaemia-specific methylation. RASSF6 was methylated in 94% (48/51) B-ALL and 41% (12/29) T-ALL, whilst RASSF10 was methylated in 16% (8/51) B-ALL and 88% (23/26) T-ALL. RASSF6 and RASSF10 expression inversely correlated with methylation which was restored by treatment with 5-aza-2\u2032deoxycytidine (5azaDC). Conclusion: This study shows the hypermethylation profile of RASSF genes in leukaemias is distinct from that of solid tumours and represents the first report of inactivation of RASSF6 or RASSF10 in cancer. These data show epigenetic inactivation of the candidate TSGs RASSF6 and RASSF10 is an extremely frequent event in the pathogenesis of childhood leukaemia. This study also warrants further investigation of the newly identified RASSF member RASSF10 and its potential role in leukaemia