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recombinant DNA technology

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https://www.readbyqxmd.com/read/28213165/hbvcircle-a-novel-tool-to-investigate-hepatitis-b-virus-covalently-closed-circular-dna
#1
Zhipeng Yan, Jing Zeng, Youjun Yu, Kunlun Xiang, Hui Hu, Xue Zhou, Lili Gu, Li Wang, Jie Zhao, John A T Young, Lu Gao
BACKGROUND & AIMS: Hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) persists as a stable episome in infected hepatocytes and serves as a template for the transcription of all viral genes. Due to the narrow host range of HBV, the development of a robust mouse model that supports cccDNA-dependent viral replication is a key hurdle in the development of novel HBV therapeutics. This study aimed to develop a novel tool to investigate HBV cccDNA. METHODS: Through minicircle technology, HBVcircle, a recombinant cccDNA, was easily generated and extracted from a genetically engineered E...
February 14, 2017: Journal of Hepatology
https://www.readbyqxmd.com/read/28212457/papillomaviruses-a-systematic-review
#2
Rodrigo Pinheiro Araldi, Suely Muro Reis Assaf, Rodrigo Franco de Carvalho, Márcio Augusto Caldas Rocha de Carvalho, Jacqueline Mazzuchelli de Souza, Roberta Fiusa Magnelli, Diego Grando Módolo, Franco Peppino Roperto, Rita de Cassia Stocco, Willy Beçak
In the last decades, a group of viruses has received great attention due to its relationship with cancer development and its wide distribution throughout the vertebrates: the papillomaviruses. In this article, we aim to review some of the most relevant reports concerning the use of bovines as an experimental model for studies related to papillomaviruses. Moreover, the obtained data contributes to the development of strategies against the clinical consequences of bovine papillomaviruses (BPV) that have led to drastic hazards to the herds...
February 16, 2017: Genetics and Molecular Biology
https://www.readbyqxmd.com/read/28209186/identification-of-immune-protective-genes-of-eimeria-maxima-through-cdna-expression-library-screening
#3
XinChao Yang, MengHui Li, JianHua Liu, YiHong Ji, XiangRui Li, LiXin Xu, RuoFeng Yan, XiaoKai Song
BACKGROUND: Eimeria maxima is one of the most prevalent Eimeria species causing avian coccidiosis, and results in huge economic loss to the global poultry industry. Current control strategies, such as anti-coccidial medication and live vaccines have been limited because of their drawbacks. The third generation anticoccidial vaccines including the recombinant vaccines as well as DNA vaccines have been suggested as a promising alternative strategy. To date, only a few protective antigens of E...
February 16, 2017: Parasites & Vectors
https://www.readbyqxmd.com/read/28191922/synergistic-integration-of-experimental-and-simulation-approaches-for-the-de-novo-design-of-silk-based-materials
#4
Wenwen Huang, Davoud Ebrahimi, Nina Dinjaski, Anna Tarakanova, Markus J Buehler, Joyce Y Wong, David L Kaplan
Tailored biomaterials with tunable functional properties are crucial for a variety of task-specific applications ranging from healthcare to sustainable, novel bio-nanodevices. To generate polymeric materials with predictive functional outcomes, exploiting designs from nature while morphing them toward non-natural systems offers an important strategy. Silks are Nature's building blocks and are produced by arthropods for a variety of uses that are essential for their survival. Due to the genetic control of encoded protein sequence, mechanical properties, biocompatibility, and biodegradability, silk proteins have been selected as prototype models to emulate for the tunable designs of biomaterial systems...
February 13, 2017: Accounts of Chemical Research
https://www.readbyqxmd.com/read/28186591/-application-of-recombinant-gp%C3%A2-a-combined-luminex-beads-for-the-detection-of-hpa-1a-antibody
#5
Sudan Tao, Ying Liu, Yanming He, Yanling Ying, Ji He, Faming Zhu
OBJECTIVE: To generate recombinant GPⅢa as an alternative source for HPA-1a antigen and combine it with Luminex xMAP beads for the detection of HPA-1a-specific alloantibody. METHODS: The full coding region of ITGB3 gene was amplified and ligated with pcDNA3.1. The recombinant plasmid was transfected into CHO cells, and those with stable expression were screened with G418. Expressed protein was identified and coupled with Luminex xMAP beads, which were then reacted with sera samples...
February 10, 2017: Zhonghua Yi Xue Yi Chuan Xue za Zhi, Zhonghua Yixue Yichuanxue Zazhi, Chinese Journal of Medical Genetics
https://www.readbyqxmd.com/read/28167937/antiviral-defenses-in-plants-through-genome-editing
#6
REVIEW
Gustavo Romay, Claude Bragard
Plant-virus interactions based-studies have contributed to increase our understanding on plant resistance mechanisms, providing new tools for crop improvement. In the last two decades, RNA interference, a post-transcriptional gene silencing approach, has been used to induce antiviral defenses in plants with the help of genetic engineering technologies. More recently, the new genome editing systems (GES) are revolutionizing the scope of tools available to confer virus resistance in plants. The most explored GES are zinc finger nucleases, transcription activator-like effector nucleases, and clustered regularly interspaced short palindromic repeats/Cas9 endonuclease...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28165224/programmable-dna-guided-artificial-restriction-enzymes
#7
Behnam Enghiad, Huimin Zhao
Restriction enzymes are essential tools for recombinant DNA technology that have revolutionized modern biological research. However, they have limited sequence specificity and availability. Here we report a Pyrococcus furiosus Argonaute (PfAgo) based platform for generating artificial restriction enzymes (AREs) capable of recognizing and cleaving DNA sequences at virtually any arbitrary site and generating defined sticky ends of varying length. Short DNA guides are used to direct PfAgo to target sites for cleavage at high temperatures (>87 °C) followed by reannealing of the cleaved single stranded DNAs...
February 6, 2017: ACS Synthetic Biology
https://www.readbyqxmd.com/read/28163952/unintended-targeting-of-dmp1-cre-reveals-a-critical-role-for-bmpr1a-signaling-in-the-gastrointestinal-mesenchyme-of-adult-mice
#8
Joohyun Lim, Joseph Burclaff, Guangxu He, Jason C Mills, Fanxin Long
Cre/loxP technology has been widely used to study cell type-specific functions of genes. Proper interpretation of such data critically depends on a clear understanding of the tissue specificity of Cre expression. The Dmp1-Cre mouse, expressing Cre from a 14-kb DNA fragment of the mouse Dmp1 gene, has become a common tool for studying gene function in osteocytes, but the presumed cell specificity is yet to be fully established. By using the Ai9 reporter line that expresses a red fluorescent protein upon Cre recombination, we find that in 2-month-old mice, Dmp1-Cre targets not only osteocytes within the bone matrix but also osteoblasts on the bone surface and preosteoblasts at the metaphyseal chondro-osseous junction...
2017: Bone Research
https://www.readbyqxmd.com/read/28154939/a-practical-guide-to-recombineering-in-photorhabdus-and-xenorhabdus
#9
Jia Yin, Hailong Wang, Ruijuan Li, Vinothkannan Ravichandran, Xiaoying Bian, Aiying Li, Qiang Tu, A Francis Stewart, Jun Fu, Youming Zhang
Fluent genetic manipulation of prokaryote genomes is still limited to only a few commonly used hosts. Ideally the advanced technologies available for cloning into recombinant Escherichia coli should also be applicable in other prokaryotes. In particular, 'recombineering' is mediated by the lambda Red operon that permits fluent and precise engineering of the E. coli genome and associated recombinant DNA. The major limitation is that host-specific phage-derived recombination systems are also required in more distant species...
February 3, 2017: Current Topics in Microbiology and Immunology
https://www.readbyqxmd.com/read/28121526/for-autoimmune-ills-biologics-bring-promise-and-problems
#10
Katherine T Adams, Peter Wehrwein
The development of recombinant DNA and other technologies has added a new dimension to care. These medications have revolutionized the treatment of rheumatoid arthritis and many of the other 80 or so autoimmune diseases. But they can be budget busters and have a tricky side effect profile.
July 2016: Managed Care
https://www.readbyqxmd.com/read/28116699/pelmo-an-optimised-in-house-cloning-vector
#11
Andrea E Ramos, Marina Muñoz, Darwin A Moreno-Pérez, Manuel A Patarroyo
DNA cloning is an essential tool regarding DNA recombinant technology as it allows the replication of foreign DNA fragments within a cell. pELMO was here constructed as an in-house cloning vector for rapid and low-cost PCR product propagation; it is an optimally designed vector containing the ccdB killer gene from the pDONR 221 plasmid, cloned into the pUC18 vector's multiple cloning site (Thermo Scientific). The ccdB killer gene has a cleavage site (CCC/GGG) for the SmaI restriction enzyme which is used for vector linearisation and cloning blunt-ended products...
December 2017: AMB Express
https://www.readbyqxmd.com/read/28116631/translational-initiatives-in-thrombolytic-therapy
#12
REVIEW
Melvin E Klegerman
Once thrombi have formed as part of the pathology defining myocardial infarction, ischemic stroke, peripheral arterial disease, deep venous thrombosis or other embolic disorders, the only clinically meaningful thrombolytic agents available for reversing the thrombogenic process are various plasminogen activators. These agents are enzymes that reverse fibrin polymerization underlying the coagulation process by converting endogenous plasminogen to plasmin, which cleaves the fibrin network to form increasingly smaller protein fragments, a process known as fibrinolysis...
January 23, 2017: Frontiers of Medicine
https://www.readbyqxmd.com/read/28114101/crispr-technology-reveals-rad-51-ical-mechanisms-of-repair-in-roundworms-an-educational-primer-for-use-with-promotion-of-homologous-recombination-by-sws-1-in-complex-with-rad-51-paralogs-in-caenorhabditis-elegans
#13
Carolyn A Turcotte, Nicolas P Andrews, Solomon A Sloat, Paula M Checchi
The mechanisms cells use to maintain genetic fidelity via DNA repair and the accuracy of these processes have garnered interest from scientists engaged in basic research to clinicians seeking improved treatment for cancer patients. Despite the continued advances, many details of DNA repair are still incompletely understood. In addition, the inherent complexity of DNA repair processes, even at the most fundamental level, makes it a challenging topic. This primer is meant to assist both educators and students in using a recent paper, "Promotion of homologous recombination by SWS-1 in complex with RAD-51 paralogs in Caenorhabditis elegans," to understand mechanisms of DNA repair...
November 2016: Genetics
https://www.readbyqxmd.com/read/28110250/detection-and-discrimination-of-maintenance-and-de-novo-cpg-methylation-events-using-methylbreak
#14
William Hsu, Augustus T Mercado, George Hsiao, Jui-Ming Yeh, Chung-Yung Chen
Understanding the principles governing the establishment and maintenance activities of DNA methyltransferases (DNMTs) can help in the development of predictive biomarkers associated with genetic disorders and diseases. A detection system was developed that distinguishes and quantifies methylation events using methylation-sensitive endonucleases and molecular beacon technology. MethylBreak (MB) is a 22-mer oligonucleotide with one hemimethylated and two unmethylated CpG sites, which are also recognition sites for Sau96I and SacII, and is attached to a fluorophore and a quencher...
January 14, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28092459/toward-high-throughput-and-multiplexed-imaging-of-genome-organization
#15
Eric F Joyce
Dr. Eric Joyce from the Department of Genetics at the University of Pennsylvania was awarded The President's Innovation award at the annual Society of Biomolecular Imaging and Informatics meeting held in Boston, September 2016. Chromosome interactions are a fundamental aspect of nuclear organization that can activate and silence genes or even direct chromosome rearrangements. However, the molecular mechanisms underlying how chromosomal segments find each other and form stable interactions within cells remain unknown...
January 2017: Assay and Drug Development Technologies
https://www.readbyqxmd.com/read/28067217/characterization-of-the-interplay-between-dna-repair-and-crispr-cas9-induced-dna-lesions-at-an-endogenous-locus
#16
Anne Bothmer, Tanushree Phadke, Luis A Barrera, Carrie M Margulies, Christina S Lee, Frank Buquicchio, Sean Moss, Hayat S Abdulkerim, William Selleck, Hariharan Jayaram, Vic E Myer, Cecilia Cotta-Ramusino
The CRISPR-Cas9 system provides a versatile toolkit for genome engineering that can introduce various DNA lesions at specific genomic locations. However, a better understanding of the nature of these lesions and the repair pathways engaged is critical to realizing the full potential of this technology. Here we characterize the different lesions arising from each Cas9 variant and the resulting repair pathway engagement. We demonstrate that the presence and polarity of the overhang structure is a critical determinant of double-strand break repair pathway choice...
January 9, 2017: Nature Communications
https://www.readbyqxmd.com/read/28060945/examining-sources-of-error-in-pcr-by-single-molecule-sequencing
#17
Vladimir Potapov, Jennifer L Ong
Next-generation sequencing technology has enabled the detection of rare genetic or somatic mutations and contributed to our understanding of disease progression and evolution. However, many next-generation sequencing technologies first rely on DNA amplification, via the Polymerase Chain Reaction (PCR), as part of sample preparation workflows. Mistakes made during PCR appear in sequencing data and contribute to false mutations that can ultimately confound genetic analysis. In this report, a single-molecule sequencing assay was used to comprehensively catalog the different types of errors introduced during PCR, including polymerase misincorporation, structure-induced template-switching, PCR-mediated recombination and DNA damage...
2017: PloS One
https://www.readbyqxmd.com/read/28045956/recombinase-polymerase-amplification-assay-a-simple-fast-and-cost-effective-alternative-to-real-time-pcr-for-specific-detection-of-feline-herpesvirus-1
#18
Jianchang Wang, Libing Liu, Jinfeng Wang, Xiaoxia Sun, Wanzhe Yuan
Feline herpesvirus 1 (FHV-1), an enveloped dsDNA virus, is one of the major pathogens of feline upper respiratory tract disease (URTD) and ocular disease. Currently, polymerase chain reaction (PCR) remains the gold standard diagnostic tool for FHV-1 infection but is relatively expensive, requires well-equipped laboratories and is not suitable for field tests. Recombinase polymerase amplification (RPA), an isothermal gene amplification technology, has been explored for the molecular diagnosis of infectious diseases...
2017: PloS One
https://www.readbyqxmd.com/read/28042089/paf53-is-essential-in-mammalian-cells-crispr-cas9-fails-to-eliminate-paf53-expression
#19
Lawrence I Rothblum, Katrina Rothblum, Eugenie Chang
When mammalian cells are nutrient and/or growth factor deprived, exposed to inhibitors of protein synthesis, stressed by heat shock or grown to confluence, rDNA transcription is essentially shut off. Various mechanisms are available to accomplish this downshift in ribosome biogenesis. Muramatsu's laboratory (Hanada et al., 1996) first demonstrated that mammalian PAF53 was essential for specific rDNA transcription and that PAF53 levels were regulated in response to growth factors. While S. cerevisae A49, the homologue of vertebrate PAF53, is not essential for viability (Liljelund et al...
December 29, 2016: Gene
https://www.readbyqxmd.com/read/28038868/marek-s-disease-vaccines-current-status-and-strategies-for-improvement-and-development-of-vector-vaccines
#20
Sanjay M Reddy, Yoshihiro Izumiya, Blanca Lupiani
Marek's disease (MD) is a lymphoproliferative viral disease of chickens, which has been controlled through vaccination since 1969. MD vaccines protect against tumors but do not provide sterilizing immunity, and thus it is generally believed that their use has contributed to increase virulence of field strains with the ability to cause MD in vaccinated chickens. Traditional methods of developing vaccines, like cell culture attenuation, have proved unsuccessful for the development of improved vaccines to protect against highly virulent MD virus (MDV) field strains...
November 24, 2016: Veterinary Microbiology
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