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recombinant DNA technology

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https://www.readbyqxmd.com/read/28540703/a-novel-platform-for-high-throughput-gene-synthesis-to-maximize-recombinant-expression-in-escherichia-coli
#1
Ana Filipa Sequeira, Joana L A Brás, Vânia O Fernandes, Catarina I P D Guerreiro, Renaud Vincentelli, Carlos M G A Fontes
Gene synthesis is becoming an important tool in many fields of recombinant DNA technology, including recombinant protein production. De novo gene synthesis is quickly replacing the classical cloning and mutagenesis procedures and allows generating nucleic acids for which no template is available. Here, we describe a high-throughput platform to design and produce multiple synthetic genes (<500 bp) for recombinant expression in Escherichia coli. This pipeline includes an innovative codon optimization algorithm that designs DNA sequences to maximize heterologous protein production in different hosts...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28526454/generation-and-characterization-of-caprine-chymosin-in-corn-seed
#2
Wen-Guo Liu, Yun-Peng Wang, Zhi-Jun Zhang, Min Wang, Qing-Xue Lv, Hong-Wei Liu, Ling-Cong Meng, Ming Lu
Chymosin is widely used in the dairy industry, and much is produced through recombinant DNA in organisms such as bacteria and tobacco. In this study, we used a new transgenic method to express caprine chymosin in corn seeds with lower cost and better storage capability. The recombinant chymosin protein was successfully expressed at an average level of 0.37 mg/g dry weight, which is 0.27% of the total soluble protein in the corn seed. Prochymosin can be activated to produce a chymosin protein with the ability to induce clotting in milk, similar to the commercial protein...
May 16, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28522247/potential-of-multi-component-antigens-for-tuberculosis-diagnosis
#3
Sana Khurshid, Madeeha Afzal, Ruqqya Khalid, M Waheed Akhtar, Mahmood Husain Qazi
Tuberculosis is one of the top ten causes of deaths worldwide. The cause of tuberculosis is a bacterium, Mycobacterium tuberculosis, which has been surviving for centuries. Immunological tests based on detecting the presence of antibodies in the sera of active TB patients against various antigens of M. tuberculosis are useful for diagnosis of TB and offer simple, rapid and cost effective methods most suitable for poor and developing countries. Several recombinant antigens have been reported so far with varying sensitivity individually, yet none had shown sensitivity higher enough to be used in a commercial test...
May 15, 2017: Biologicals: Journal of the International Association of Biological Standardization
https://www.readbyqxmd.com/read/28508691/corifollitropin-alfa-compared-with-follitropin-beta-in-gnrh-antagonist-ovarian-stimulation-protocols-in-an-unselected-population-undergoing-ivf-icsi
#4
Charalampos Siristatidis, Konstantinos Dafopoulos, Nikolas Christoforidis, George Anifandis, Vasileios Pergialiotis, Nikolaos Papantoniou
Recombinant DNA technologies have produced Corifollitropin alfa (CFa) used during IVF/ICSI in order to keep the circulating FSH levels above the threshold necessary to support multi-follicular growth for a week. In this prospective case-control study, we compared 70 participants treated with 150 μg CFa combined with 150 IU of follitropin beta (study group) with 70 subfertile participants with matching baseline characteristics, conforming with the same inclusion criteria and treated with an antagonist protocol using follitropin beta (control group)...
May 16, 2017: Gynecological Endocrinology
https://www.readbyqxmd.com/read/28495315/accelerated-mass-production-of-influenza-virus-seed-stocks-in-hek-293-suspension-cell-cultures-by-reverse-genetics
#5
Ernest Milián, Thomas Julien, Rafael Biaggio, Alina Venereo-Sanchez, Johnny Montes, Aziza P Manceur, Sven Ansorge, Emma Petiot, Manuel Rosa-Calatrava, Amine Kamen
Despite major advances in developing capacities and alternative technologies to egg-based production of influenza vaccines, responsiveness to an influenza pandemic threat is limited by the time it takes to generate a Candidate Vaccine Virus (CVV) as reported by the 2015 WHO Informal Consultation report titled "Influenza Vaccine Response during the Start of a Pandemic". In previous work, we have shown that HEK-293 cell culture in suspension and serum free medium is an efficient production platform for cell culture manufacturing of influenza candidate vaccines...
May 8, 2017: Vaccine
https://www.readbyqxmd.com/read/28494568/-the-research-advances-and-applications-of-genome-editing-in-hereditary-eye-diseases
#6
S W Cai, Y Zhang, M Z Hou, Y Liu, X R Li
Genome editing is a cutting-edge technology that generates DNA double strand breaks at the specific genomic DNA sequence through nuclease recognition and cleavage, and then achieves insertion, replacement, or deletion of the target gene via endogenous DNA repair mechanisms, such as non-homologous end joining, homology directed repair, and homologous recombination. So far, more than 600 human hereditary eye diseases and systemic hereditary diseases with ocular phenotypes have been found. However, most of these diseases are of incompletely elucidated pathogenesis and without effective therapies...
May 11, 2017: [Zhonghua Yan Ke za Zhi] Chinese Journal of Ophthalmology
https://www.readbyqxmd.com/read/28487820/recombinant-dna-technology-and-click-chemistry-a-powerful-combination-for-generating-a-hybrid-elastin-like-statherin-hydrogel-to-control-calcium-phosphate-mineralization
#7
Mohamed Hamed Misbah, Mercedes Santos, Luis Quintanilla, Christina Günter, Matilde Alonso, Andreas Taubert, José Carlos Rodríguez-Cabello
Understanding the mechanisms responsible for generating different phases and morphologies of calcium phosphate by elastin-like recombinamers is supreme for bioengineering of advanced multifunctional materials. The generation of such multifunctional hybrid materials depends on the properties of their counterparts and the way in which they are assembled. The success of this assembly depends on the different approaches used, such as recombinant DNA technology and click chemistry. In the present work, an elastin-like recombinamer bearing lysine amino acids distributed along the recombinamer chain has been cross-linked via Huisgen [2 + 3] cycloaddition...
2017: Beilstein Journal of Nanotechnology
https://www.readbyqxmd.com/read/28485577/site-selective-conjugation-of-native-proteins-with-dna
#8
Julie B Trads, Thomas Tørring, Kurt V Gothelf
Conjugation of DNA to proteins is increasingly used in academia and industry to provide proteins with tags for identification or handles for hybridization to other DNA strands. Assay technologies such as immuno-PCR and proximity ligation and the imaging technology DNA-PAINT require DNA-protein conjugates. In DNA nanotechnology, the DNA handle is exploited to precisely position proteins by self-assembly. For these applications, site-selective conjugation is almost always desired because fully functional proteins are required to maintain the specificity of antibodies and the activity of enzymes...
May 9, 2017: Accounts of Chemical Research
https://www.readbyqxmd.com/read/28479163/cell-penetrating-peptides-cpps-from-delivery-of-nucleic-acids-and-antigens-to-transduction-of-engineered-nucleases-for-application-in-transgenesis
#9
REVIEW
Gandhi Rádis-Baptista, Iana S Campelo, Jean-Étienne R L Morlighem, Luciana M Melo, Vicente J F Freitas
Cell-penetrating peptides (CPPs) have been studied for their capacity to translocate across the lipid membrane of several cell types. In membrane translocation, these peptides can remarkably transport biologically active hydrophilic molecules, such as pharmaceuticals, nucleic acids (DNA and RNA) and even high-molecular-weight proteins, Fig. 3 into the cell cytoplasm and organelles. The development of CPPs as transduction agents includes the modification of gene and protein expression, the reprogramming and differentiation of induced pluripotent stem cells and the preparation of cellular vaccines...
May 4, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28458052/production-of-monoclonal-antibodies-and-development-of-a-quantitative-immuno-polymerase-chain-reaction-assay-to-detect-and-quantify-recombinant-glutathione-s-transferase
#10
Abud Je, Luque Eh, Ramos Jg, Rodriguez Ha
GST-tagged proteins are important tools for the production of recombinant proteins. Removal of GST tag from its fusion protein, frequently by harsh chemical treatments or proteolytic methods, is often required. Thus, the monitoring of the proteins in tag-free form requires a significant effort to determine the remnants of GST during purification process. In the present study, we developed both a conventional enzyme-linked immunosorbent assay (ELISA) and an immuno-polymerase chain reaction (IPCR) assay, both specific for detection of recombinant GST (rGST)...
April 27, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28433722/high-precision-genome-sequencing-of-engineered-gluconobacter-oxydans-621h-by-combining-long-nanopore-and-short-accurate-illumina-reads
#11
Angela Kranz, Alexander Vogel, Ursula Degner, Ines Kiefler, Michael Bott, Björn Usadel, Tino Polen
State of the art and novel high-throughput DNA sequencing technologies enable fascinating opportunities and applications in the life sciences including microbial genomics. Short high-quality read data already enable not only microbial genome sequencing, yet can be inadequately to solve problems in genome assemblies and for the analysis of structural variants, especially in engineered microbial cell factories. Single-molecule real-time sequencing technologies generating long reads promise to solve such assembly problems...
April 19, 2017: Journal of Biotechnology
https://www.readbyqxmd.com/read/28429327/large-scale-cultivation-of-euglena
#12
Kengo Suzuki
From the middle of the twentieth century, microalgae have been exploited as a candidate biomass source of food and other products. One such candidate source is the fast-proliferating microalga Euglena gracilis. The commercial cultivation of E. gracilis began in 2007, after the success of its outdoor mass cultivation and improvement of the harvesting and drying methods suitable for Euglena cells. The commercialization of Euglena production is based on the strategy of "5Fs of Biomass," which refers to the development and production of commercial products including food, fiber, feed, fertilizer, and fuel from biomass...
2017: Advances in Experimental Medicine and Biology
https://www.readbyqxmd.com/read/28414239/a-novel-sample-preparation-for-shotgun-proteomics-characterization-of-hcps-in-antibodies
#13
Lihua Huang, Ning Wang, Charles E Mitchell, Tammy Brownlee, Steven R Maple, Michael R De Felippis
Residual host cell proteins (HCPs) in biopharmaceuticals derived from recombinant DNA technology can present potential safety risks to patients or compromise product stability. Thus, the downstream purification process is designed to demonstrate robust removal of these impurities. ELISA using polyclonal anti-HCP antibodies as reagents for capture, detection, and quantitation purposes is most commonly used to monitor HCP removal during process development, but this technique has limitations. More recently, LC-MS for residual HCP characterization has emerged as a powerful tool to support purification process development...
May 16, 2017: Analytical Chemistry
https://www.readbyqxmd.com/read/28394093/genome-editing-technologies-to-improve-research-reproduction-and-production-in-pigs
#14
REVIEW
Kevin D Wells, Randall S Prather
The ability to directly manipulate the pig genome through genetic engineering has been available to the research community for over three decades. This technology has progressed from the random insertion of foreign DNA, via a variety of techniques (pronuclear microinjection, sperm mediated gene transfer, and integration of mobile genetic elements), to manipulation of endogenous genes, via homologous recombination in somatic cells followed by somatic cell nuclear transfer. Over the last few years, designer nucleases facilitated the development of techniques that provide efficient ways to introduce foreign DNA or to modify endogenous genes in eggs, zygotes, or somatic cells...
April 10, 2017: Molecular Reproduction and Development
https://www.readbyqxmd.com/read/28390488/functional-fusion-proteins-and-prevention-of-electrode-fouling-for-a-sensitive-electrochemical-immunosensor
#15
A-Ram Kim, Tae Jung Park, Minseok S Kim, In-Ho Kim, Ki-Suk Kim, Kwang Hoe Chung, Sungho Ko
A highly sensitive electrochemical immunosensor was developed by preventing electrode fouling and using a novel fusion protein of silica binding polypeptides (SBP)-protein G (ProG) created by recombinant DNA technology as a functional crosslinker for rapid and self-oriented immobilization of antibodies onto silica nanoparticles (SiNPs). Antibody immobilization onto the SiNPs by the SBP-ProG could rapidly be achieved without any chemical treatment. The immunosensor was fabricated through bonding of a partially gold-deposited cyclic olefin copolymer (COC) (top substrate) and gold patterned interdigitated array COC electrode (bottom substrate)...
May 15, 2017: Analytica Chimica Acta
https://www.readbyqxmd.com/read/28387174/institutional-oversight-of-occupational-health-and-safety-for-research-programs-involving-biohazards
#16
Melissa C Dyson, Calvin B Carpenter, Lesley A Colby
Research with hazardous biologic materials (biohazards) is essential to the progress of medicine and science. The fieldof microbiology has rapidly advanced over the years, partially due to the development of new scientific methods such as recombinant DNA technology, synthetic biology, viral vectors, and the use of genetically modified animals. This research poses a potential risk to personnel as well as the public and the environment. Institutions must have appropriate oversight and take appropriate steps to mitigate the risks of working with these biologic hazards...
April 6, 2017: Comparative Medicine
https://www.readbyqxmd.com/read/28369428/novel-approaches-to-neisseria-meningitidis-vaccine-design
#17
Myron Christodoulides, John Heckels
A range of vaccines is available for preventing life-threatening diseases caused by infection with Neisseria meningitidis (meningococcus, Men). Capsule polysaccharide (CPS)-conjugate vaccines are successful prophylactics for serogroup MenA, MenC, MenW and MenY infections and outer membrane vesicle (OMV) vaccines have been used successfully for controlling clonal serogroup MenB infections. MenB vaccines based on recombinant proteins identified by reverse vaccinology (Bexsero®) and proteomics (Trumenba®) approaches have recently been licensed and Bexsero® has been introduced into the UK infant immunization program...
March 22, 2017: Pathogens and Disease
https://www.readbyqxmd.com/read/28361829/genetic-characterization-of-human-herpesvirus-type-1-full-length-genome-sequence-of-strain-obtained-from-an-encephalitis-case-from-india
#18
Vijay P Bondre, Vasudha Sankararaman, Vijaysinh Andhare, Manisha Tupekar, Gajanan N Sapkal
BACKGROUND & OBJECTIVES: Human herpes simplex virus 1 (HSV-1) is the most common cause of sporadic encephalitis in humans that contributes to >10 per cent of the encephalitis cases occurring worldwide. Availability of limited full genome sequences from a small number of isolates resulted in poor understanding of host and viral factors responsible for variable clinical outcome. In this study genetic relationship, extent and source of recombination using full-length genome sequence derived from a newly isolated HSV-1 isolate was studied in comparison with those sampled from patients with varied clinical outcome...
November 2016: Indian Journal of Medical Research
https://www.readbyqxmd.com/read/28360231/combination-of-short-read-long-read-and-optical-mapping-assemblies-reveals-large-scale-tandem-repeat-arrays-with-population-genetic-implications
#19
Matthias H Weissensteiner, Andy W C Pang, Ignas Bunikis, Ida Höijer, Olga Vinnere-Petterson, Alexander Suh, Jochen B W Wolf
Accurate and contiguous genome assembly is key to a comprehensive understanding of the processes shaping genomic diversity and evolution. Yet, it is frequently constrained by constitutive heterochromatin, usually characterized by highly repetitive DNA. As a key feature of genome architecture associated with centromeric and subtelomeric regions, it locally influences meiotic recombination. In this study, we assess the impact of large tandem repeat arrays on the recombination rate landscape in an avian speciation model, the Eurasian crow...
May 2017: Genome Research
https://www.readbyqxmd.com/read/28325290/rna-guided-crispr-cas9-system-mediated-engineering-of-acute-myeloid-leukemia-mutations
#20
Oliver Brabetz, Vijay Alla, Linus Angenendt, Christoph Schliemann, Wolfgang E Berdel, Maria-Francisca Arteaga, Jan-Henrik Mikesch
Current acute myeloid leukemia (AML) disease models face severe limitations because most of them induce un-physiological gene expressions that do not represent conditions in AML patients and/or depend on external promoters for regulation of gene expression/repression. Furthermore, many AML models are based on reciprocal chromosomal translocations that only reflect the minority of AML patients, whereas more than 50% of patients have a normal karyotype. The majority of AML, however, is driven by somatic mutations...
March 17, 2017: Molecular Therapy. Nucleic Acids
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