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recombinant DNA technology

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https://www.readbyqxmd.com/read/28634346/generating-conditional-gene-knockouts-in-plasmodium-a-toolkit-to-produce-stable-dicre-recombinase-expressing-parasite-lines-using-crispr-cas9
#1
Ellen Knuepfer, Marta Napiorkowska, Christiaan van Ooij, Anthony A Holder
Successful establishment of CRISPR/Cas9 genome editing technology in Plasmodium spp. has provided a powerful tool to transform Plasmodium falciparum into a genetically more tractable organism. Conditional gene regulation approaches are required to study the function of gene products critical for growth and erythrocyte invasion of blood stage parasites. Here we employ CRISPR/Cas9 to facilitate use of the dimerisable Cre-recombinase (DiCre) that is frequently used to mediate the excision and loss of loxP-flanked DNA sequences in a rapamycin controlled manner...
June 20, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28622634/type-of-gonadotropin-used-during-controlled-ovarian-stimulation-induces-differential-gene-expression-in-human-cumulus-cells-a-randomized-study
#2
María Cruz, Antonio Requena, David Agudo, Juan Antonio García-Velasco
BACKGROUND: The cumulus-oocyte complex plays a central role in the regulation of folliculogenesis where it is important for the maturation, reprogramming, and fertilization of oocytes. Consequently, cumulus cell gene expression profiling is being explored as a promising method for assessing oocyte competence in the near future. Through DNA microarray technology, we analyzed the potential differences in the gene expression profiles of cumulus cells from preovulatory follicles after controlled ovarian stimulation using different types of gonadotropins...
June 8, 2017: European Journal of Obstetrics, Gynecology, and Reproductive Biology
https://www.readbyqxmd.com/read/28612264/application-of-recombinant-hemagglutinin-proteins-as-alternative-antigen-standards-for-pandemic-influenza-vaccines
#3
Yejin Choi, Seong Yi Kwon, Ho Jung Oh, Sunbo Shim, Seokkee Chang, Hye Joo Chung, Do Keun Kim, Younsang Park, Younghee Lee
OBJECTIVES: The single radial immunodiffusion (SRID) assay, used to quantify hemagglutinin (HA) in influenza vaccines, requires reference reagents; however, because centralized production of reference reagents may slow the emergency deployment of vaccines, alternatives are needed. RESULTS: We investigated the production of HA proteins using recombinant DNA technology, rather than a traditional egg-based production process. The HA proteins were then used in an SRID assay as a reference antigen...
June 13, 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/28602961/progress-and-biotechnological-prospects-in-fish-transgenesis
#4
REVIEW
Fernanda M P Tonelli, Samyra M S N Lacerda, Flávia C P Tonelli, Guilherme M J Costa, Luiz Renato de França, Rodrigo R Resende
The history of transgenesis is marked by milestones such as the development of cellular transdifferentiation, recombinant DNA, genetic modification of target cells, and finally, the generation of simpler genetically modified organisms (e.g. bacteria and mice). The first transgenic fish was developed in 1984, and since then, continuing technological advancements to improve gene transfer have led to more rapid, accurate, and efficient generation of transgenic animals. Among the established methods are microinjection, electroporation, lipofection, viral vectors, and gene targeting...
June 8, 2017: Biotechnology Advances
https://www.readbyqxmd.com/read/28599921/molecular-advances-in-microbial-aminopeptidases
#5
REVIEW
Arya Nandan, Kesavan Madhavan Nampoothiri
Aminopeptidases are exopeptidases that catalyze the hydrolysis of amino acid residues from the N terminus of peptides and proteins. They are widely and diversely used for protein hydrolysis in industrial and research applications. They form a large enzyme family in microorganisms and most of the sequenced microbial genomes contain several aminopeptidase coding genes. Various approaches are being used to enhance the yield and desired properties of these enzymes to make it more suited for industrial applications...
May 19, 2017: Bioresource Technology
https://www.readbyqxmd.com/read/28596938/genome-edited-t-cell-therapies
#6
REVIEW
Juliette M K M Delhove, Waseem Qasim
PURPOSE OF REVIEW: Alternative approaches to conventional drug-based cancer treatments have seen T cell therapies deployed more widely over the last decade. This is largely due to their ability to target and kill specific cell types based on receptor recognition. Introduction of recombinant T cell receptors (TCRs) using viral vectors and HLA-independent T cell therapies using chimeric antigen receptors (CARs) are discussed. This article reviews the tools used for genome editing, with particular emphasis on the applications of site-specific DNA nuclease mediated editing for T cell therapies...
2017: Current Stem Cell Reports
https://www.readbyqxmd.com/read/28592991/aav-based-dual-reporter-circuit-for-monitoring-cell-signaling-in-living-human-cells
#7
Zhiwen Zhang, Zachary Stickney, Natalie Duong, Kevin Curley, Biao Lu
BACKGROUND: High-throughput methods based on molecular reporters have greatly advanced our knowledge of cell signaling in mammalian cells. However, their ability to monitor various types of cells is markedly limited by the inefficiency of reporter gene delivery. Recombinant adeno-associated virus (AAV) vectors are efficient tools widely used for delivering and expressing transgenes in diverse animal cells in vitro and in vivo. Here we present the design, construction and validation of a novel AAV-based dual-reporter circuit that can be used to monitor and quantify cell signaling in living human cells...
2017: Journal of Biological Engineering
https://www.readbyqxmd.com/read/28592330/state-of-play-and-clinical-prospects-of-antibody-gene-transfer
#8
REVIEW
Kevin Hollevoet, Paul J Declerck
Recombinant monoclonal antibodies (mAbs) are one of today's most successful therapeutic classes in inflammatory diseases and oncology. A wider accessibility and implementation, however, is hampered by the high product cost and prolonged need for frequent administration. The surge in more effective mAb combination therapies further adds to the costs and risk of toxicity. To address these issues, antibody gene transfer seeks to administer to patients the mAb-encoding nucleotide sequence, rather than the mAb protein...
June 7, 2017: Journal of Translational Medicine
https://www.readbyqxmd.com/read/28581731/synthesis-of-a-bifunctional-peptide-inhibitor-igg1-fc-fusion-that-suppresses-experimental-autoimmune-encephalomyelitis
#9
Derek R White, Zahra Khedri, Paul Kiptoo, Teruna J Siahaan, Thomas J Tolbert
Multiple sclerosis (MS) is a neurodegenerative disease that is estimated to affect over 2.3 million people worldwide. The exact cause for this disease is unknown but involves immune system attack and destruction of the myelin protein surrounding the neurons in the central nervous system. One promising class of compounds that selectively prevent the activation of immune cells involved in the pathway leading to myelin destruction are Bifunctional Peptide Inhibitors (BPIs). Treatment with BPIs reduces neurodegenerative symptoms in Experimental Autoimmune Encephalomyelitis (EAE), a mouse model of MS...
June 5, 2017: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/28565744/-chromosomal-large-fragment-deletion-induced-by-crispr-cas9-gene-editing-system
#10
L H Cheng, Y Liu, T Niu
Objective: Using CRISPR-Cas9 gene editing technology to achieve a number of genes co-deletion on the same chromosome. Methods: CRISPR-Cas9 lentiviral plasmid that could induce deletion of Aloxe3-Alox12b-Alox8 cluster genes located on mouse 11B3 chromosome was constructed via molecular clone. HEK293T cells were transfected to package lentivirus of CRISPR or Cas9 cDNA, then mouse NIH3T3 cells were infected by lentivirus and genomic DNA of these cells was extracted. The deleted fragment was amplified by PCR, TA clone, Sanger sequencing and other techniques were used to confirm the deletion of Aloxe3-Alox12b-Alox8 cluster genes...
May 14, 2017: Zhonghua Xue Ye Xue za Zhi, Zhonghua Xueyexue Zazhi
https://www.readbyqxmd.com/read/28559484/finding-the-needle-in-the-haystack-the-use-of-microfluidic-droplet-technology-to-identify-vitamin-secreting-lactic-acid-bacteria
#11
Jun Chen, Mike Vestergaard, Thomas Glasdam Jensen, Jing Shen, Martin Dufva, Christian Solem, Peter Ruhdal Jensen
Efficient screening technologies aim to reduce both the time and the cost required for identifying rare mutants possessing a phenotype of interest in a mutagenized population. In this study, we combined a mild mutagenesis strategy with high-throughput screening based on microfluidic droplet technology to identify Lactococcus lactis variants secreting vitamin B2 (riboflavin). Initially, we used a roseoflavin-resistant mutant of L. lactis strain MG1363, JC017, which secreted low levels of riboflavin. By using fluorescence-activated droplet sorting, several mutants that secreted riboflavin more efficiently than JC017 were readily isolated from the mutagenesis library...
May 30, 2017: MBio
https://www.readbyqxmd.com/read/28558726/development-and-application-of-triple-antibody-sandwich-enzyme-linked-immunosorbent-assays-for-begomovirus-detection-using-monoclonal-antibodies-against-tomato-yellow-leaf-curl-thailand-virus
#12
Channarong Seepiban, Saengsoon Charoenvilaisiri, Nuchnard Warin, Anjana Bhunchoth, Namthip Phironrit, Bencharong Phuangrat, Orawan Chatchawankanphanich, Supat Attathom, Oraprapai Gajanandana
BACKGROUND: Tomato yellow leaf curl Thailand virus, TYLCTHV, is a begomovirus that causes severe losses of tomato crops in Thailand as well as several countries in Southeast and East Asia. The development of monoclonal antibodies (MAbs) and serological methods for detecting TYLCTHV is essential for epidemiological studies and screening for virus-resistant cultivars. METHODS: The recombinant coat protein (CP) of TYLCTHV was expressed in Escherichia coli and used to generate MAbs against TYLCTHV through hybridoma technology...
May 30, 2017: Virology Journal
https://www.readbyqxmd.com/read/28548882/finding-conducting-and-nurturing-science-a-virologist-s-memoir
#13
Anna Marie Ann Skalka Skalka
My laboratory investigations have been driven by an abiding interest in understanding the consequences of genetic rearrangement in evolution and disease, and in using viruses to elucidate fundamental mechanisms in biology. Starting with bacteriophages and moving to the retroviruses, my use of the tools of genetics, molecular biology, biochemistry, and biophysics has spanned more than half a century-from the time when DNA structure was just discovered to the present day of big data and epigenetics. Both riding and contributing to the successive waves of technology, my laboratory has elucidated fundamental mechanisms in DNA replication, repair, and recombination...
May 26, 2017: Annual Review of Virology
https://www.readbyqxmd.com/read/28540703/a-novel-platform-for-high-throughput-gene-synthesis-to-maximize-recombinant-expression-in-escherichia-coli
#14
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
#15
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 17, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28522247/potential-of-multi-component-antigens-for-tuberculosis-diagnosis
#16
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
#17
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
#18
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
#19
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
#20
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
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