This study aimed to clone the phytoene synthase (psy) gene from beta-carotene-rich sweet potato as an alternative gene for genetic transformation of cassava. The researchers cloned the full psy open reading frame from sweet potato using rapid amplification of cDNA ends. They obtained a 1029bp open reading frame encoding a 342 amino acid protein with high similarity to tomato psy. The researchers plan to express this psy gene in cassava under a root-specific promoter to increase beta-carotene levels and verify that it produces a functional enzyme for carotenoid biosynthesis.
kbDNA Inc. offers recombinant dna products for research and development. Review kbDNA's quality Recombinant DNA products at https://www.kbdna.com/recombinants
kbDNA Inc. offers recombinant dna products for research and development. Review kbDNA's quality Recombinant DNA products at https://www.kbdna.com/recombinants
Cloning and expression of the Nodamura virus RNA-dependent RNA polymerase
Poster presentation at Society for the Advancement of Chicanos and Native Americans in Science (SACNAS) National Conference, October 2012, Seatltle, WA
The GeneArt® Gene Synthesis service consists of chemical synthesis, cloning, and sequence verification of virtually any desired genetic sequence. You will receive a bacterial stab and/or purified plasmid containing your synthesized gene—ready for downstream applications.
Whether you have limited cloning experience or simply want to save time, the GeneArt® Gene Synthesis service helps you move your ideas from the planning stage to the laboratory more quickly. Benefit from our experience in successfully producing over 180,000 constructs for customers as diverse as large pharmaceutical companies, biotechnology start-ups, and basic research institutions. The comparison shown in the figure below highlights the time and effort saved compared to traditional cloning. For more information visit:
https://www.invitrogen.com/site/us/en/home/Products-and-Services/Applications/Cloning/gene-synthesis.html?CID=genesynthesis-SS-12312
The analysis of all transcripts within a cell is of essential importance. Molecular biology provides many approaches to clone RNA transcripts into cDNA. Large cDNA collections are in the public domain to serve the research community. Today, however, new high-speed sequencing methods allow a much deeper view into transcriptomes than possible by classical cloning.
Enabling RNA-Seq With Limited RNA Using Whole Transcriptome AmplificationQIAGEN
RNA-Seq was developed to perform transcriptome profiling and provides a highly precise measurement of expression levels of transcripts and their isoforms. Normally, RNA-Seq analysis requires at least 500 ng –1 μg of total RNA. When working with small biopsies, single cells (such as circulating tumor cells), or other limited material, whole transcriptome amplification (WTA) is normally required. Various WTA methods overcome limited RNA availability and enable transcriptome analysis from limited material or even single cells. In standard PCR-based WTA procedures, however, bias from uneven coverage of cDNA regions with high GC or AT content or amplification errors can lead to the loss of transcripts and wrong variant calling. Here, we compare a standard RNA-Seq library preparation method and the REPLI-g RNA library protocol. The REPLI-g procedure is a PCR-free protocol to efficiently generate RNA-Seq libraries from small amounts of RNA or a single cell in 6.5–7 hours. The REPLI-g protocol uses whole transcriptome amplification based on multiple displacement amplification (MDA), combined with an efficient library adaptor ligation procedure, to prepare RNA-Seq libraries from small RNA amounts. The procedure demonstrates high fidelity, minimal bias and retention of sample‘s transcriptional profile. Compared to standard RNA-Seq library prep, the REPLI-g protocol demonstrates similar reproducibility and sensitivity in transcript detection.
AGRF in conjunction with EMBL Australia recently organised a workshop at Monash University Clayton. This workshop was targeted at beginners and biologists who are new to analysing Next-Gen Sequencing data. The workshop also aimed to provide users with a snapshot of bioinformatics and data analysis tips on how to begin to analyse project data. An introduction to RNA-seq data analysis was presented by AGRF Senior Bioinformatician Dr. Sonika Tyagi.
Presented: 1st August 2012
Durante la Semana de la Agricultura y la Alimentación, el Programa de Investigación del CGIAR en Cambio Climático, Agricultura y Seguridad Alimentaria – CCAFS, la Organización de las Naciones Unidas para la Alimentación y la Agricultura, FAO, y el Centro Internacional de Agricultura Tropical – CIAT, apoyaron la II Reunión Internacional de Ministros y altas autoridades de agricultura sobre agricultura sostenible y cambio climático con un documento base y su presentación sobre los retos que representa el cambio climático para la agricultura en Latino América y el Caribe.
Taller sobre intervenciones en nutrición, género y agricultura: situación actual y oportunidades futuras’, organizado por el CIAT y HarvestPlus en Ciudad de Guatemala. Leer más: http://ow.ly/XNIv30mGYBv
More Related Content
Similar to Poster71: A phytoene synthase (psy) open reading frame form B-carotene-rich sweet potato as an alterntative gene for cassava genetic trasformation
Cloning and expression of the Nodamura virus RNA-dependent RNA polymerase
Poster presentation at Society for the Advancement of Chicanos and Native Americans in Science (SACNAS) National Conference, October 2012, Seatltle, WA
The GeneArt® Gene Synthesis service consists of chemical synthesis, cloning, and sequence verification of virtually any desired genetic sequence. You will receive a bacterial stab and/or purified plasmid containing your synthesized gene—ready for downstream applications.
Whether you have limited cloning experience or simply want to save time, the GeneArt® Gene Synthesis service helps you move your ideas from the planning stage to the laboratory more quickly. Benefit from our experience in successfully producing over 180,000 constructs for customers as diverse as large pharmaceutical companies, biotechnology start-ups, and basic research institutions. The comparison shown in the figure below highlights the time and effort saved compared to traditional cloning. For more information visit:
https://www.invitrogen.com/site/us/en/home/Products-and-Services/Applications/Cloning/gene-synthesis.html?CID=genesynthesis-SS-12312
The analysis of all transcripts within a cell is of essential importance. Molecular biology provides many approaches to clone RNA transcripts into cDNA. Large cDNA collections are in the public domain to serve the research community. Today, however, new high-speed sequencing methods allow a much deeper view into transcriptomes than possible by classical cloning.
Enabling RNA-Seq With Limited RNA Using Whole Transcriptome AmplificationQIAGEN
RNA-Seq was developed to perform transcriptome profiling and provides a highly precise measurement of expression levels of transcripts and their isoforms. Normally, RNA-Seq analysis requires at least 500 ng –1 μg of total RNA. When working with small biopsies, single cells (such as circulating tumor cells), or other limited material, whole transcriptome amplification (WTA) is normally required. Various WTA methods overcome limited RNA availability and enable transcriptome analysis from limited material or even single cells. In standard PCR-based WTA procedures, however, bias from uneven coverage of cDNA regions with high GC or AT content or amplification errors can lead to the loss of transcripts and wrong variant calling. Here, we compare a standard RNA-Seq library preparation method and the REPLI-g RNA library protocol. The REPLI-g procedure is a PCR-free protocol to efficiently generate RNA-Seq libraries from small amounts of RNA or a single cell in 6.5–7 hours. The REPLI-g protocol uses whole transcriptome amplification based on multiple displacement amplification (MDA), combined with an efficient library adaptor ligation procedure, to prepare RNA-Seq libraries from small RNA amounts. The procedure demonstrates high fidelity, minimal bias and retention of sample‘s transcriptional profile. Compared to standard RNA-Seq library prep, the REPLI-g protocol demonstrates similar reproducibility and sensitivity in transcript detection.
AGRF in conjunction with EMBL Australia recently organised a workshop at Monash University Clayton. This workshop was targeted at beginners and biologists who are new to analysing Next-Gen Sequencing data. The workshop also aimed to provide users with a snapshot of bioinformatics and data analysis tips on how to begin to analyse project data. An introduction to RNA-seq data analysis was presented by AGRF Senior Bioinformatician Dr. Sonika Tyagi.
Presented: 1st August 2012
Sequencing the transcriptome reveals complex layers of regulation, Department...Copenhagenomics
Sequencing the transcriptome reveals complex layers of regulation
Similar to Poster71: A phytoene synthase (psy) open reading frame form B-carotene-rich sweet potato as an alterntative gene for cassava genetic trasformation (20)
Durante la Semana de la Agricultura y la Alimentación, el Programa de Investigación del CGIAR en Cambio Climático, Agricultura y Seguridad Alimentaria – CCAFS, la Organización de las Naciones Unidas para la Alimentación y la Agricultura, FAO, y el Centro Internacional de Agricultura Tropical – CIAT, apoyaron la II Reunión Internacional de Ministros y altas autoridades de agricultura sobre agricultura sostenible y cambio climático con un documento base y su presentación sobre los retos que representa el cambio climático para la agricultura en Latino América y el Caribe.
Taller sobre intervenciones en nutrición, género y agricultura: situación actual y oportunidades futuras’, organizado por el CIAT y HarvestPlus en Ciudad de Guatemala. Leer más: http://ow.ly/XNIv30mGYBv
Impacto de las intervenciones agricolas y de salud para reducir la deficienci...CIAT
Taller sobre intervenciones en nutrición, género y agricultura: situación actual y oportunidades futuras’, organizado por el CIAT y HarvestPlus en Ciudad de Guatemala. Leer más: http://ow.ly/XNIv30mGYBv.
Presentado por Byron Reyes, CIAT/ Harvestplus
Agricultura sensible a la nutrición en el Altiplano. Explorando las perspecti...CIAT
Taller sobre intervenciones en nutrición, género y agricultura: situación actual y oportunidades futuras’, organizado por el CIAT y HarvestPlus en Ciudad de Guatemala. Leer más: http://ow.ly/XNIv30mGYBv
El rol de los padres en la nutrición del hogarCIAT
Taller sobre intervenciones en nutrición, género y agricultura: situación actual y oportunidades futuras’, organizado por el CIAT y HarvestPlus en Ciudad de Guatemala. Leer más: http://ow.ly/XNIv30mGYBv
Jennifer Twyman, Líder de investigación de Género en el CIAT
Scaling up soil carbon enhancement contributing to mitigate climate changeCIAT
The 4 per 1000 Africa Symposium - Building synergies across Africa to advance on soils for food security and climate, Johannesburg, South Africa 24-26 October 2018
Rolf Sommer, Kristin Piikki, Mats Söderström, Sylvia Nyawira, Mayesse da Silva, Wuletawu Abera and
Job Kihara
Impacto del Cambio Climático en la Agricultura de República DominicanaCIAT
El Banco Interamericano de Desarrollo (BID) y el Centro Internacional de Agricultura Tropical (CIAT), con el apoyo de los Programas de Investigación de CGIAR sobre Políticas, Instituciones y Mercados (PIM) y sobre Cambio Climático, Agricultura y Seguridad Alimentaria (CCAFS), se han asociado para comprender, a través de la ciencia, el impacto del cambio climático en cultivos claves y el impacto económico en la productividad de la agricultura en países de ALC.
BioTerra: Nuevo sistema de monitoreo de la biodiversidad en desarrollo por el...CIAT
BioTerra es un sistema innovador de monitoreo de la biodiversidad y sus amenazas desarrollado por el Programa Riqueza Natural de la Agencia de los Estados Unidos para el Desarrollo Internacional (USAID), y sus socios locales – el Centro Internacional de Agricultura Tropical (CIAT) y el Instituto Alexander von Humboldt (IAvH) – para apoyar al gobierno colombiano en el cumplimiento de las metas y compromisos de conservación de la biodiversidad. Este sistema busca complementar y aunar esfuerzos existentes de monitoreo de la biodiversidad y sus amenazas, a nivel nacional y regional.
Cacao for Peace Activities for Tackling the Cadmium in Cacao Issue in Colo...CIAT
El taller ‘Cacao libre de cadmio’, organizado por el CIAT, CIRAD, y la AFD, se lleva a cabo del 12 al 14 de marzo en la sede del CIAT en Palmira,y tiene como objetivo integrar un consorcio de actores y disciplinas claves de la región, así como elaborar un proyecto de investigación aplicada que dé respuesta a este problema que afecta a los cacaoteros de Colombia, Perú y Ecuador. http://ow.ly/J43p30iU0UZ
Tackling cadmium in cacao and derived products – from farm to forkCIAT
El taller ‘Cacao libre de cadmio’, organizado por el CIAT, CIRAD, y la AFD, se lleva a cabo del 12 al 14 de marzo en la sede del CIAT en Palmira,y tiene como objetivo integrar un consorcio de actores y disciplinas claves de la región, así como elaborar un proyecto de investigación aplicada que dé respuesta a este problema que afecta a los cacaoteros de Colombia, Perú y Ecuador. http://ow.ly/J43p30iU0UZ
Cadmium bioaccumulation and gastric bioaccessibility in cacao: A field study ...CIAT
El taller ‘Cacao libre de cadmio’, organizado por el CIAT, CIRAD, y la AFD, se lleva a cabo del 12 al 14 de marzo en la sede del CIAT en Palmira,y tiene como objetivo integrar un consorcio de actores y disciplinas claves de la región, así como elaborar un proyecto de investigación aplicada que dé respuesta a este problema que afecta a los cacaoteros de Colombia, Perú y Ecuador. http://ow.ly/J43p30iU0UZ
Geographical Information System Mapping for Optimized Cacao Production in Col...CIAT
El taller ‘Cacao libre de cadmio’, organizado por el CIAT, CIRAD, y la AFD, se lleva a cabo del 12 al 14 de marzo en la sede del CIAT en Palmira,y tiene como objetivo integrar un consorcio de actores y disciplinas claves de la región, así como elaborar un proyecto de investigación aplicada que dé respuesta a este problema que afecta a los cacaoteros de Colombia, Perú y Ecuador. http://ow.ly/J43p30iU0UZ
El taller ‘Cacao libre de cadmio’, organizado por el CIAT, CIRAD, y la AFD, se lleva a cabo del 12 al 14 de marzo en la sede del CIAT en Palmira,y tiene como objetivo integrar un consorcio de actores y disciplinas claves de la región, así como elaborar un proyecto de investigación aplicada que dé respuesta a este problema que afecta a los cacaoteros de Colombia, Perú y Ecuador. http://ow.ly/J43p30iU0UZ
Técnicas para disminuir la disponibilidad de cadmio en suelos de cacaoterasCIAT
El taller ‘Cacao libre de cadmio’, organizado por el CIAT, CIRAD, y la AFD, se lleva a cabo del 12 al 14 de marzo en la sede del CIAT en Palmira,y tiene como objetivo integrar un consorcio de actores y disciplinas claves de la región, así como elaborar un proyecto de investigación aplicada que dé respuesta a este problema que afecta a los cacaoteros de Colombia, Perú y Ecuador. http://ow.ly/J43p30iU0UZ
El taller ‘Cacao libre de cadmio’, organizado por el CIAT, CIRAD, y la AFD, se lleva a cabo del 12 al 14 de marzo en la sede del CIAT en Palmira,y tiene como objetivo integrar un consorcio de actores y disciplinas claves de la región, así como elaborar un proyecto de investigación aplicada que dé respuesta a este problema que afecta a los cacaoteros de Colombia, Perú y Ecuador. http://ow.ly/J43p30iU0UZ
El taller ‘Cacao libre de cadmio’, organizado por el CIAT, CIRAD, y la AFD, se lleva a cabo del 12 al 14 de marzo en la sede del CIAT en Palmira,y tiene como objetivo integrar un consorcio de actores y disciplinas claves de la región, así como elaborar un proyecto de investigación aplicada que dé respuesta a este problema que afecta a los cacaoteros de Colombia, Perú y Ecuador. http://ow.ly/J43p30iU0UZ
de la región, así como elaborar un proyecto de investigación aplicada que dé respuesta a este problema que afecta a los cacaoteros de Colombia, Perú y Ecuador. http://ow.ly/J43p30iU0UZ
El taller ‘Cacao libre de cadmio’, organizado por el CIAT, CIRAD, y la AFD, se lleva a cabo del 12 al 14 de marzo en la sede del CIAT en Palmira,y tiene como objetivo integrar un consorcio de actores y disciplinas claves de la región, así como elaborar un proyecto de investigación aplicada que dé respuesta a este problema que afecta a los cacaoteros de Colombia, Perú y Ecuador. http://ow.ly/J43p30iU0UZ
Smart TV Buyer Insights Survey 2024 by 91mobiles.pdf91mobiles
91mobiles recently conducted a Smart TV Buyer Insights Survey in which we asked over 3,000 respondents about the TV they own, aspects they look at on a new TV, and their TV buying preferences.
UiPath Test Automation using UiPath Test Suite series, part 4DianaGray10
Welcome to UiPath Test Automation using UiPath Test Suite series part 4. In this session, we will cover Test Manager overview along with SAP heatmap.
The UiPath Test Manager overview with SAP heatmap webinar offers a concise yet comprehensive exploration of the role of a Test Manager within SAP environments, coupled with the utilization of heatmaps for effective testing strategies.
Participants will gain insights into the responsibilities, challenges, and best practices associated with test management in SAP projects. Additionally, the webinar delves into the significance of heatmaps as a visual aid for identifying testing priorities, areas of risk, and resource allocation within SAP landscapes. Through this session, attendees can expect to enhance their understanding of test management principles while learning practical approaches to optimize testing processes in SAP environments using heatmap visualization techniques
What will you get from this session?
1. Insights into SAP testing best practices
2. Heatmap utilization for testing
3. Optimization of testing processes
4. Demo
Topics covered:
Execution from the test manager
Orchestrator execution result
Defect reporting
SAP heatmap example with demo
Speaker:
Deepak Rai, Automation Practice Lead, Boundaryless Group and UiPath MVP
GDG Cloud Southlake #33: Boule & Rebala: Effective AppSec in SDLC using Deplo...James Anderson
Effective Application Security in Software Delivery lifecycle using Deployment Firewall and DBOM
The modern software delivery process (or the CI/CD process) includes many tools, distributed teams, open-source code, and cloud platforms. Constant focus on speed to release software to market, along with the traditional slow and manual security checks has caused gaps in continuous security as an important piece in the software supply chain. Today organizations feel more susceptible to external and internal cyber threats due to the vast attack surface in their applications supply chain and the lack of end-to-end governance and risk management.
The software team must secure its software delivery process to avoid vulnerability and security breaches. This needs to be achieved with existing tool chains and without extensive rework of the delivery processes. This talk will present strategies and techniques for providing visibility into the true risk of the existing vulnerabilities, preventing the introduction of security issues in the software, resolving vulnerabilities in production environments quickly, and capturing the deployment bill of materials (DBOM).
Speakers:
Bob Boule
Robert Boule is a technology enthusiast with PASSION for technology and making things work along with a knack for helping others understand how things work. He comes with around 20 years of solution engineering experience in application security, software continuous delivery, and SaaS platforms. He is known for his dynamic presentations in CI/CD and application security integrated in software delivery lifecycle.
Gopinath Rebala
Gopinath Rebala is the CTO of OpsMx, where he has overall responsibility for the machine learning and data processing architectures for Secure Software Delivery. Gopi also has a strong connection with our customers, leading design and architecture for strategic implementations. Gopi is a frequent speaker and well-known leader in continuous delivery and integrating security into software delivery.
SAP Sapphire 2024 - ASUG301 building better apps with SAP Fiori.pdfPeter Spielvogel
Building better applications for business users with SAP Fiori.
• What is SAP Fiori and why it matters to you
• How a better user experience drives measurable business benefits
• How to get started with SAP Fiori today
• How SAP Fiori elements accelerates application development
• How SAP Build Code includes SAP Fiori tools and other generative artificial intelligence capabilities
• How SAP Fiori paves the way for using AI in SAP apps
GraphSummit Singapore | The Art of the Possible with Graph - Q2 2024Neo4j
Neha Bajwa, Vice President of Product Marketing, Neo4j
Join us as we explore breakthrough innovations enabled by interconnected data and AI. Discover firsthand how organizations use relationships in data to uncover contextual insights and solve our most pressing challenges – from optimizing supply chains, detecting fraud, and improving customer experiences to accelerating drug discoveries.
Threats to mobile devices are more prevalent and increasing in scope and complexity. Users of mobile devices desire to take full advantage of the features
available on those devices, but many of the features provide convenience and capability but sacrifice security. This best practices guide outlines steps the users can take to better protect personal devices and information.
PHP Frameworks: I want to break free (IPC Berlin 2024)Ralf Eggert
In this presentation, we examine the challenges and limitations of relying too heavily on PHP frameworks in web development. We discuss the history of PHP and its frameworks to understand how this dependence has evolved. The focus will be on providing concrete tips and strategies to reduce reliance on these frameworks, based on real-world examples and practical considerations. The goal is to equip developers with the skills and knowledge to create more flexible and future-proof web applications. We'll explore the importance of maintaining autonomy in a rapidly changing tech landscape and how to make informed decisions in PHP development.
This talk is aimed at encouraging a more independent approach to using PHP frameworks, moving towards a more flexible and future-proof approach to PHP development.
Sudheer Mechineni, Head of Application Frameworks, Standard Chartered Bank
Discover how Standard Chartered Bank harnessed the power of Neo4j to transform complex data access challenges into a dynamic, scalable graph database solution. This keynote will cover their journey from initial adoption to deploying a fully automated, enterprise-grade causal cluster, highlighting key strategies for modelling organisational changes and ensuring robust disaster recovery. Learn how these innovations have not only enhanced Standard Chartered Bank’s data infrastructure but also positioned them as pioneers in the banking sector’s adoption of graph technology.
GraphSummit Singapore | The Future of Agility: Supercharging Digital Transfor...Neo4j
Leonard Jayamohan, Partner & Generative AI Lead, Deloitte
This keynote will reveal how Deloitte leverages Neo4j’s graph power for groundbreaking digital twin solutions, achieving a staggering 100x performance boost. Discover the essential role knowledge graphs play in successful generative AI implementations. Plus, get an exclusive look at an innovative Neo4j + Generative AI solution Deloitte is developing in-house.
Dr. Sean Tan, Head of Data Science, Changi Airport Group
Discover how Changi Airport Group (CAG) leverages graph technologies and generative AI to revolutionize their search capabilities. This session delves into the unique search needs of CAG’s diverse passengers and customers, showcasing how graph data structures enhance the accuracy and relevance of AI-generated search results, mitigating the risk of “hallucinations” and improving the overall customer journey.
GraphRAG is All You need? LLM & Knowledge GraphGuy Korland
Guy Korland, CEO and Co-founder of FalkorDB, will review two articles on the integration of language models with knowledge graphs.
1. Unifying Large Language Models and Knowledge Graphs: A Roadmap.
https://arxiv.org/abs/2306.08302
2. Microsoft Research's GraphRAG paper and a review paper on various uses of knowledge graphs:
https://www.microsoft.com/en-us/research/blog/graphrag-unlocking-llm-discovery-on-narrative-private-data/
Climate Impact of Software Testing at Nordic Testing DaysKari Kakkonen
My slides at Nordic Testing Days 6.6.2024
Climate impact / sustainability of software testing discussed on the talk. ICT and testing must carry their part of global responsibility to help with the climat warming. We can minimize the carbon footprint but we can also have a carbon handprint, a positive impact on the climate. Quality characteristics can be added with sustainability, and then measured continuously. Test environments can be used less, and in smaller scale and on demand. Test techniques can be used in optimizing or minimizing number of tests. Test automation can be used to speed up testing.
GridMate - End to end testing is a critical piece to ensure quality and avoid...ThomasParaiso2
End to end testing is a critical piece to ensure quality and avoid regressions. In this session, we share our journey building an E2E testing pipeline for GridMate components (LWC and Aura) using Cypress, JSForce, FakerJS…
DevOps and Testing slides at DASA ConnectKari Kakkonen
My and Rik Marselis slides at 30.5.2024 DASA Connect conference. We discuss about what is testing, then what is agile testing and finally what is Testing in DevOps. Finally we had lovely workshop with the participants trying to find out different ways to think about quality and testing in different parts of the DevOps infinity loop.
Removing Uninteresting Bytes in Software FuzzingAftab Hussain
Imagine a world where software fuzzing, the process of mutating bytes in test seeds to uncover hidden and erroneous program behaviors, becomes faster and more effective. A lot depends on the initial seeds, which can significantly dictate the trajectory of a fuzzing campaign, particularly in terms of how long it takes to uncover interesting behaviour in your code. We introduce DIAR, a technique designed to speedup fuzzing campaigns by pinpointing and eliminating those uninteresting bytes in the seeds. Picture this: instead of wasting valuable resources on meaningless mutations in large, bloated seeds, DIAR removes the unnecessary bytes, streamlining the entire process.
In this work, we equipped AFL, a popular fuzzer, with DIAR and examined two critical Linux libraries -- Libxml's xmllint, a tool for parsing xml documents, and Binutil's readelf, an essential debugging and security analysis command-line tool used to display detailed information about ELF (Executable and Linkable Format). Our preliminary results show that AFL+DIAR does not only discover new paths more quickly but also achieves higher coverage overall. This work thus showcases how starting with lean and optimized seeds can lead to faster, more comprehensive fuzzing campaigns -- and DIAR helps you find such seeds.
- These are slides of the talk given at IEEE International Conference on Software Testing Verification and Validation Workshop, ICSTW 2022.
FIDO Alliance Osaka Seminar: The WebAuthn API and Discoverable Credentials.pdf
Poster71: A phytoene synthase (psy) open reading frame form B-carotene-rich sweet potato as an alterntative gene for cassava genetic trasformation
1. A phytoene synthase (psy) open reading frame from β-carotene-rich sweet potato as an
alternative gene for cassava genetic transformation.
Parra S1, Toro N2, Beltran JA1, Chavarriaga P 1, and Tohme J1
1. Conservation and Use of Tropical Genetic Resources, CIAT, AA 6713 , Cali, Colombia.
2. Universidad del Valle, Departamento de Biología, Cali, Colombia.
INTRODUCTION • 5’ RACE
By means of genetic transformation we seek to increase the β-carotene
content in cassava roots by over-expressing, in a tissue-specific With the first amplification towrads the cDNA 5’ end we obtained a
manner, genes of the carotenoid synthesis pathway. Currently we are band of approximately 800bp (Figure 2A). It was isolated and
using the bacterial gene crtB as a source of Phytoene Synthase (psy), sequenced. The 783bp obtained was similar (E-value = 0) to the C.
which has proven to be efficient to enhance carotenoid production in canephora psy sequence. The comparison between the sequence
fruits, seeds and roots of several crops obtained and other psy sequence showed a lack of a transit peptide.
Therefore we performed a second amplification towards the cDNA
However, plant derived genes, besides having the advantage of being 5’end and obtained an extra 200bp (Figure 2B) product. After
better accepted by the public, may be a more efficient source of PSY, characterizing the product, 143 nucleotides were confirmed to be
as it has been proven with Golden Rice. We are following the same psy sequence, with similarity (E-value = 6.3x10-23) to the L.
approach for cassava and therefore have cloned the psy gene sequence esculentum psy gene.
from β-carotene rich sweet potato storage roots (Ipomoea batatas) by
means of rapid amplification of cDNA ends (RACE).
METHODS 0.8Kb
0.2Kb
A B
• Primers design
Figure 2. Amplification results to the 5’cDNA end. A. First amplification.
B. Second amplification.
Degenerate forward primers were designed considering the alignment
result of psy sequences from Coffea canephora (Robusta Coffee),
Gentiana lutea (Gentian), Lycium barbarum (Goji), Capsicum annuum With the three different sequences of the cDNA ends we predicted a
(Pepper) and Nicotiana langsdorffii (Tobacco), to amplify the 3’ cDNA contig and suggested an open reading frame (ORF) for the sweet
end. With the known sequences obtained after 3’ end amplification potato psy gene.
and characterization, we designed specific reverse primers to
synthesize the cDNA and amplify the 5’ end.
• Complete psy ORF isolation
• Sequence isolation Using a pair of specific primers we isolated the complete psy ORF.
The amplification product had 1000bp (Figure 3A) approximately,
Single strand cDNA with
which was isolated and characterized. We obtained a 1029bp open
adapter ccc reading frame that has a predicted protein of 342 aminoacids. The
comparison with the Swissprot database showed high similarity (E-
value = 3.9x10-176) with the enzyme PSY of L. esculentum (Figure
3B).
Specific primers design
5’cDNA sequence
Amplification with outer
primers.
3’cDNA sequence Specific primers
design
1Kb
PCR
A B
Nucleotide
Figure 3. Sweet potato psy ORF. A. Amplification result with specific primers. B.
sequence Alignment result between the predicted aminoacids sequences and others reported in
Nested amplification with the Swissprot database.
inner primers
RESULTS AND DISCUSSION PERSPECTIVES
• 3’ RACE
• Cloning the psy ORF in an expression vector under a cassava root-
A product of approximately 500bp (Fig 1) was obtained with the specific promoter, or a constitutive promoter, to transform cassava
degenerate primers in the 3’cDNA end nested amplification. Then, embryogenic cells, to verify that the ORF produces a functional
this product was isolated from the gel and sequenced. The 480bp enzyme.
sequence was similar (E-value = 4.8x10-88) to the Lycopersicon
esculentum (tomato) psy gene. • Cloning the promoter of the psy sequence to use it as an alternative
source of regulatory regions for the expression of carotenogenic genes
in cassava roots.
0.5Kb
Figure 1. Nested amplification result to the 3’cDNA end
Date prepared: July 2008