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Molecular Biotechnology

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https://read.qxmd.com/read/30747382/biochemical-kinetic-and-computational-structural-characterization-of-shikimate-kinase-from-methicillin-resistant-staphylococcus-aureus
#1
Alejandro Favela-Candia, Alfredo Téllez-Valencia, Mara Campos-Almazán, Erick Sierra-Campos, Mónica Valdez-Solana, Jesús Oria-Hernández, Adriana Castillo-Villanueva, Hugo Nájera, Claudia Avitia-Domínguez
One of the most widespread pathogens worldwide is methicillin-resistant Staphylococcus aureus, a bacterium that provokes severe life-threatening illnesses both in hospitals and in the community. The principal challenge lies in the resistance of MRSA to current treatments, which encourages the study of different molecular targets that could be used to develop new drugs against this infectious agent. With this goal, a detailed characterization of shikimate kinase from this microorganism (SaSK) is described. The results showed that SaSK has a Km of 0...
February 12, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30747381/correction-to-production-of-a-recombinant-dermaseptin-peptide-in-nicotiana-tabacum-hairy-roots-with-enhanced-antimicrobial-activity
#2
Marzieh Varasteh Shams, Farhad Nazarian-Firouzabadi, Ahmad Ismaili, Reza Shirzadian-Khorramabad
The original version of this article unfortunately contained a mistake in the unit "μg/l". The unit "μg/l" should be corrected to "μg/ml" throughout the paper.
February 12, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30734200/responsiveness-and-adaptation-to-salt-stress-of-the-redox-responsive-transcription-factor-1-rrtf1-gene-are-controlled-by-its-promoter
#3
Elham R S Soliman, Peter Meyer
The REDOX-RESPONSIVE TRANSCRIPTION FACTOR 1 (RRTF1) gene encodes a member of the ERF/AP2 transcription factor family involved in redox homeostasis. The RRTF1 gene shows tissue-specific responsiveness to various abiotic stress treatments including a response to salt stress in roots. An interesting feature of this response is an adaptation phase that follows its activation, when promoter levels revert to a base line level, even if salt stress is maintained. It is unclear if adaption is controlled by a switch in promoter activity or by changes in transcript levels...
February 7, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30729436/analysis-of-in-vivo-activity-of-the-bovine-cholesterol-hydroxylase-lyase-system-proteins-expressed-in-escherichia-coli
#4
V S Efimova, L V Isaeva, M A Rubtsov, L A Novikova
The cholesterol hydroxylase/lyase (CHL) system, located in the mitochondria of the mammalian adrenal cortex cells, consists of cytochrome P450scc (CYP11A1), adrenodoxin (Adx), and adrenodoxin reductase (AdR) and performs the first stage of the steroidogenesis: AdR and Adx enable the electron transfer between NADPH and cytochrome P450scc, and P450scc catalyzes the conversion of cholesterol into pregnenolone. CHL system was reconstructed in Escherichia coli using the polycistronic plasmid pTrc99A/CHL. In E. coli cells, the recombinant proteins form the catalytically active system...
February 7, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30729435/phage-display-libraries-from-binders-to-targeted-drug-delivery-and-human-therapeutics
#5
REVIEW
Mouldy Sioud
The application of repertoire selection technologies for the study and characterization of tumor heterogeneity is an area of great interest in the field of tumor biology and immunotherapy. Among the most promising approaches, phage display has been successfully used to select peptides and antibody fragments to a variety of different targets, including cancer cells, immune cells and cytokines. Peptides selected from phage display have been used to guide the delivery of lytic peptides, cytotoxic drugs, and nanoparticles to cancer cells with the aim to obtain more efficient and less toxic therapeutics...
February 7, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30721405/construction-and-characterization-of-a-medium-copy-number-expression-vector-carrying-auto-inducible-dps-promoter-to-overproduce-a-bacterial-superoxide-dismutase-in-escherichia-coli
#6
Debbie Soefie Retnoningrum, I Wayan Martadi Santika, Suryanata Kesuma, Syahdu Ayu Ekowati, Catur Riani
Medium copy number expression vector and auto-inducible promoter could be a solution for producing recombinant therapeutic proteins in industrial scale regarding plasmid stability, cost, and product quality. This work aimed to construct a medium copy number pBR322-based expression vector carrying auto-inducible promoter, determine its ability to express heterologous gene, and study its segregational stability. Three stationary-phase promoters of Escherichia coli genes (gadA, dps and sbmC) were used to produce a superoxide dismutase from Staphylococcus equorum (rMnSODSeq) coding region from pBR322Δtet (pBR322-mini)...
February 5, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30661170/the-growth-differentiation-factor-11-is-involved-in-skin-fibroblast-ageing-and-is-induced-by-a-preparation-of-peptides-and-sugars-derived-from-plant-cell-cultures
#7
Annalisa Tito, Ani Barbulova, Claudia Zappelli, Marilisa Leone, Menotti Ruvo, Flavia Anna Mercurio, Angela Chambery, Rosita Russo, Maria Gabriella Colucci, Fabio Apone
Ageing is a complex and progressive phenomenon, during which the accumulation of morphological and chemical changes seriously compromises the capacity of the cells to proliferate and fulfil their biological tasks. The increase in the average age of the world population, associated with a higher occurrence of age-related diseases, is prompting scientific research to look for new strategies and molecular targets that may help in alleviating age-related phenotypes. Growth factors, responsible for modulating several aging markers in many tissues and organs, represent valuable targets to fight age-associated dysfunctions...
January 19, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30656573/validated-nuclear-based-transgene-expression-regulated-by-the-fea1-iron-responsive-promoter-in-the-green-alga-chlamydomonas-reinhardtii
#8
Paula Barjona do Nascimento Coutinho, Christine Friedl, Marcus Heilmann, Rainer Buchholz, Stephanie Christine Stute
Microalgae are in the focus for the production of recombinant proteins in research and potential commercial application. Inducible promoters represent important tools that potentially allow the expression of recombinant proteins at higher rates. In general, they are used to separate the culture growth phase from the production phase by initiating product formation after high cell densities have been achieved. This potentially offers a higher space-time yield, consequently improving the economics of a process...
January 17, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30656572/building-bridges-between-structural-and-network-based-systems-biology
#9
REVIEW
Christos T Chasapis
The integration of structural and network-based systems biology is paramount for the improved understanding of how the proteins interact to modulate the behavior of complex biological systems. This review presents the current literature on the computational studies that combine these two scientific fields focusing on two main approaches: network-based analysis of the structure and dynamics of proteins and the in silico reconstruction of protein-protein interaction (PPI) networks or expansion of existed protein interactomes driven by structural annotations...
January 17, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30649664/production-of-a-recombinant-dermaseptin-peptide-in-nicotiana-tabacum-hairy-roots-with-enhanced-antimicrobial-activity
#10
Marzieh Varasteh Shams, Farhad Nazarian-Firouzabadi, Ahmad Ismaili, Reza Shirzadian-Khorramabad
Expression of strong antimicrobial peptides in plants is of great interest to combat a wide range of plant pathogens. To bring the Dermaseptin B1 (DrsB1) peptide to the intimate contact of the plant pathogens cell wall surface, the DrsB1 encoding sequence was fused to the C-terminal part of the two copies of the chitin-binding domain (CBD) of the Avr4 effector protein and used for Agrobacterium rhizogenes-mediated transformation. The expression of the recombinant protein in the tobacco hairy roots (HRs) was confirmed by molecular analysis...
January 16, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30649663/isolation-and-expression-analysis-of-three-types-of-%C3%AE-carbonic-anhydrases-from-the-antarctic-alga-chlamydomonas-sp-ice-l-under-different-light-stress-treatments
#11
Chongli Shi, Meiling An, Jinlai Miao, Yingying He, Zhou Zheng, Changfeng Qu, Xixi Wang, Huan Lin, Junhong Liu
Carbonic anhydrases (CAs) are a class of zinc-containing metalloenzymes that can reversibly catalyse the hydration reaction of carbon dioxide. Antarctic algae are the most critical component of the Antarctic ecosystem; algae can enter the carbon cycle food chain by fixing carbon dioxide from the air. In this study, the complete open reading frames (ORFs) of CA1 (GenBank ID KY826431), CA2 (GenBank ID KY826432), and CA3 (GenBank ID KY826433), encoding CAs in the Antarctic ice microalga Chlamydomonas. sp. ICE-L, were successfully cloned using reverse transcription-polymerase chain reaction (RT-PCR)...
January 16, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30644027/crispr-cas9-mediated-multiply-targeted-mutagenesis-in-orange-and-purple-carrot-plants
#12
Zhi-Sheng Xu, Kai Feng, Ai-Sheng Xiong
The clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9) system has been successfully used for precise genome editing in many plant species, including in carrot cells, very recently. However, no stable gene-editing carrot plants were obtained with CRISPR/Cas9 system to date. In the present study, four sgRNA expression cassettes, individually driven by four different promoters and assembled in a single CRISPR/Cas9 vector, were transformed into carrots using Agrobacterium-mediated genetic transformation...
January 14, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30637606/understanding-m-6-a-function-through-uncovering-the-diversity-roles-of-yth-domain-containing-proteins
#13
REVIEW
Y L Zhao, Y H Liu, R F Wu, Z Bi, Y X Yao, Q Liu, Y Z Wang, X X Wang
N6 -methyladenosine (m6 A) is the most abundant-internal modification of eukaryotic mRNA. m6 A can be installed and removed by specific enzymes. The "writer," "eraser," and "reader" of m6 A modification have been reported. These discoveries facilitate our understanding of the functional significance of m6 A. m6 A plays an essential role in diverse biological processes by recruiting the corresponding YTH domain-containing proteins, as well as recruiting additional translation initiation factors...
January 12, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30600448/development-of-soybean-yellow-mottle-mosaic-virus-based-expression-vector-for-heterologous-protein-expression-in-french-bean
#14
Nagamani Sandra, A Abdul Kader Jailani, Rakesh Kumar Jain, Bikash Mandal
Plant virus-based vectors provide attractive and valuable tools for rapid production of recombinant protein in large quantities as they produce systemic infections in differentiated plant tissues. In the present study, we engineered the Soybean yellow mottle mosaic virus (SYMMV) as a gene expression vector which is a promising candidate for systemic expression of foreign proteins in French bean plants. Full virus vector strategy was exploited for insertion of foreign gene by inserting MCS through PCR in the circular pJET-SYMMV clone...
January 2, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30600447/ethylene-response-factor-erf-family-proteins-in-abiotic-stresses-and-crispr-cas9-genome-editing-of-erfs-for-multiple-abiotic-stress-tolerance-in-crop-plants-a-review
#15
REVIEW
Johni Debbarma, Yogita N Sarki, Banashree Saikia, Hari Prasanna Deka Boruah, Dhanawantari L Singha, Channakeshavaiah Chikkaputtaiah
Abiotic stresses such as extreme heat, cold, drought, and salt have brought alteration in plant growth and development, threatening crop yield and quality leading to global food insecurity. Many factors plays crucial role in regulating various plant growth and developmental processes during abiotic stresses. Ethylene response factors (ERFs) are AP2/ERF superfamily proteins belonging to the largest family of transcription factors known to participate during multiple abiotic stress tolerance such as salt, drought, heat, and cold with well-conserved DNA-binding domain...
January 2, 2019: Molecular Biotechnology
https://read.qxmd.com/read/30560399/improved-delivery-of-crispr-cas9-system-using-magnetic-nanoparticles-into-porcine-fibroblast
#16
Magdalena Hryhorowicz, Bartosz Grześkowiak, Natalia Mazurkiewicz, Paweł Śledziński, Daniel Lipiński, Ryszard Słomski
Genetically modified pigs play an important role in agriculture and biomedical research; hence, new efficient methods are needed to obtain genetically engineered cells and animals. The clustered regularly interspaced short palindromic repeats (CRISPR)/Cas (CRISPR-associated) system represents an effective genome editing tool. It consists of two key molecules: single guide RNA (sgRNA) and the Cas9 endonuclease that can be introduced into the cells as one plasmid. Typical delivery methods for CRISPR/Cas9 components are limited by low transfection efficiency or toxic effects on cells...
December 17, 2018: Molecular Biotechnology
https://read.qxmd.com/read/30543053/engineering-chlamydomonas-reinhardtii-for-expression-of-functionally-active-human-interferon-%C3%AE
#17
Yassin El-Ayouty, Islam El-Manawy, Sherif Nasih, Emad Hamdy, Rashad Kebeish
Human interferon (IFN) are secreted cytokines that play a major regulatory role in response to various infections. Commercially, IFN-α has been approved to treat many chronic viral diseases as well as a variety of cancers and different types of leukemia. In this study, a binary vector containing human IFN-α2a gene under the regulation of the cauliflower mosaic virus 35S promoter was constructed. IFN-œ2a expression cassette was transferred to Chlamydomonas reinhardtii cells via Agrobacterium-mediated transformation method...
December 12, 2018: Molecular Biotechnology
https://read.qxmd.com/read/30539415/new-nuclease-from-extremely-psychrophilic-microorganism-psychromonas-ingrahamii-37-identification-and-characterization
#18
Natalia Maciejewska, Rafał Walkusz, Marcin Olszewski, Aneta Szymańska
Nucleases are an important group of hydrolases that degrade nucleic acids, with broad spectrum of applications in science and industry. In this paper, we report the identification and characterization of the nuclease from extremely psychrophilic bacterium Psychromonas ingrahamii that grows exponentially at 5 °C, but may also grow at even lower temperatures (down to - 12 °C). The putative endonuclease I gene, identified in the genome of P. ingrahamii, was cloned and expressed in Pichia pastoris. The recombinant protein was purified and its nucleolytic features were studied...
December 11, 2018: Molecular Biotechnology
https://read.qxmd.com/read/30539414/generation-and-characterization-of-fibroblast-specific-basigin-knockout-mice
#19
Xiao-Dong Wu, Meng-Yao Zhang, Ya-Tong Chen, Hui Yao, Qing Zhang, Wen-Jing Wang, Da-Fu Fu, Ren-Ji Wei, Jia-Yu Zhang, Yin Li, Dan Dang, Hui-Jie Bian, Jing Xu, Zhi-Nan Chen
Basigin is a well-known extracellular stimulator of fibroblasts and may confer resistance to apoptosis of fibroblasts in vitro under some pathological status, but its exact function in fibroblasts and the underlying mechanism remain poorly understood. The systematic Basigin gene knockout leads to the perinatal lethality of mice, which limits the delineation of its function in vivo. In this study, we generated a fibroblast-specific Basigin knock-out mouse model and demonstrated the successful deletion of Basigin in fibroblasts...
December 11, 2018: Molecular Biotechnology
https://read.qxmd.com/read/30506260/expression-of-recombinant-pfceltos-antigen-in-the-chloroplast-of-chlamydomonas-reinhardtii-and-its-potential-use-in-detection-of-malaria
#20
Shabnam Shamriz, Hamideh Ofoghi
Malaria is a serious but preventable and treatable infectious disease that is found in over 100 countries around the world. Correct and rapid diagnosis of malaria infection can rescue the patient of getting sicker and reduces the risk of disease spreading among humans. Chlamydomonas reinhardtii chloroplast is an attractive platform for expressing malaria antigens because it is capable of folding complex proteins, including those requiring disulfide bond formation, while lack the ability to glycosylate proteins; a valuable quality of any malaria protein expression system, since the Plasmodium parasite lacks N-linked glycosylation machinery...
November 30, 2018: Molecular Biotechnology
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