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Molecular Biotechnology

Ge Zhao, Jianrong Wang, Qingyun Tang, Dongming Lan, Yonghua Wang
MAS1 is a lipase isolated from Streptomyces sp. strain W007 with potential application in biotechnology. Structural analysis of MAS1 lipase showed that eight amino acids with bulkier side located in the substrate-binding pocket may be involved in affecting catalytic performance. Alanine substitutions of those residues were conducted to reduce steric clash of catalyzed pocket and probe their functional roles. The k cat /K m of mutants H108A, F153A, and V233A increased to 2.3-, 2.1-, and 1.4-fold, respectively...
February 15, 2018: Molecular Biotechnology
Diego A Díaz-Dinamarca, José I Jerias, Daniel A Soto, Jorge A Soto, Natalia V Díaz, Yessica Y Leyton, Rodrigo A Villegas, Alexis M Kalergis, Abel E Vásquez
Group B Streptococcus (GBS) is the leading cause of neonatal meningitis and a common pathogen in livestock and aquaculture industries around the world. Conjugate polysaccharide and protein-based vaccines are under development. The surface immunogenic protein (SIP) is a conserved protein in all GBS serotypes and has been shown to be a good target for vaccine development. The expression of recombinant proteins in Escherichia coli cells has been shown to be useful in the development of vaccines, and the protein purification is a factor affecting their immunogenicity...
February 13, 2018: Molecular Biotechnology
Wilton Mbinda, Omwoyo Ombori, Christina Dixelius, Richard Oduor
Sweetpotato is a significant crop which is widely cultivated particularly in the developing countries with high and stable yield. However, drought stress is a major limiting factor that antagonistically influences the crop's productivity. Dehydration stress caused by drought causes aggregation of reactive oxygen species (ROS) in plants, and aldose reductases are first-line safeguards against ROS caused by oxidative stress. In the present study, we generated transgenic sweetpotato plants expressing aldose reductase, XvAld1 isolated from Xerophyta viscosa under the control of a stress-inducible promoter via Agrobacterium-mediated transformation...
February 9, 2018: Molecular Biotechnology
Chen Liang, Ziwei Liu, Chaoping Liu, Yimeng Li, Hongyu Yuan, Tianwen Wang
Classic and conventional procedures in molecular cloning are inherent compositions in modern molecular biological experiments and are frequently involved in daily laboratory activities. They take up the majority of the total time input in spite of the availability of well-designed specialized commercial kits. A similar situation is also in the field of biotechnology. Fortunately, microwave/ultrasonic irradiation has been found to be capable of speeding up these processes, such as proteolysis in sample preparation for proteomics research, and digestion, ligation, (de)phosphorylation of DNA with the corresponding enzymes, even the introduction of DNA samples to recipient cells, and biotransformation (e...
February 2, 2018: Molecular Biotechnology
Vishal Kumar, Arun Kumar Dangi, Pratyoosh Shukla
Xylanases are one of the important hydrolytic enzymes which hydrolyze the β-1, 4 xylosidic linkage of the backbone of the xylan polymeric chain which consists of xylose subunits. Xylanases are mainly found in plant cell walls and are produced by several kinds of microorganisms such as fungi, bacteria, yeast, and some protozoans. The fungi are considered as most potent xylanase producers than that of yeast and bacteria. There is a broad series of industrial applications for the thermostable xylanase as an industrial enzyme...
January 29, 2018: Molecular Biotechnology
Justin Kirke, Noah Kaplan, Stephanie Velez, Xiao-Lu Jin, Paveena Vichyavichien, Xing-Hai Zhang
A promoter is an essential structural component of a gene that controls its transcription activity in different development stages and in response to various environmental stimuli. Knowledge of promoter functionality in heterologous systems is important in the study of gene regulation and biotechnological application. In order to explore the activity of the pepper capsaicin synthase gene (PUN1) promoter, gene constructs of pPUN1::GUS (for β-glucuronidase) and pPUN1::NtKED (for a tobacco wound-responsive protein) were introduced into tobacco and tomato, respectively, and their activities were examined...
January 25, 2018: Molecular Biotechnology
Mahdi Eskandarian Boroujeni, Mossa Gardaneh
Viral gene delivery is hailed as a great milestone in gene-based therapeutic approaches. The human immunodeficiency virus-derived lentiviral vectors (LVs) are advantageous in infecting both dividing and non-dividing cells leading to continuous expression of transgenes. A variety of protocols are available for concentration of LVs. We primarily generated our internal ribosome entry site (IRES)-based LVs. Virus titration and transduction efficiency were compared between various strategies that included sucrose cushion centrifugation (SCC), protein column ultrafiltration and polyethylene glycol precipitation...
January 10, 2018: Molecular Biotechnology
Honglian Wang, Xia Zhong, Jianchun Li, Menglian Zhu, Lu Wang, Xingli Ji, Junming Fan, Li Wang
IgA protease is secreted by various mucosal pathogenic bacteria which can cleave human immunoglobulin A1 (IgA1) in its hinge region. In addition to be considered as a virulence factor, it's reported that IgA protease can also be used for IgA nephropathy (IgAN) treatment. Our previous study identified bacteria H. influenzae 49247 expressed high activity of IgA protease with promised application in IgAN therapy. In this study, we cloned the IgA protease gene of H. influenzae 49247 with degenerate primers. Alignment analysis indicated that H...
January 9, 2018: Molecular Biotechnology
Grzegorz Machnik, Estera Skudrzyk, Łukasz Bułdak, Jarosław Ruczyński, Agnieszka Kozłowska, Piotr Mucha, Piotr Rekowski, Witold Szkróbka, Marcin Basiak, Aleksandra Bołdys, Helena Sławska, Bogusław Okopień
In the presented assay, we elaborated a method for distinguishing sequences that are genetically closely related to each other. This is particularly important in a situation where a fine balance of the allele abundance is a point of research interest. We developed a peptide nucleic acid (PNA) strand invasion technique for the differentiation between multiple sclerosis-associated retrovirus (MSRV) and ERVWE1 sequences, both molecularly similar, belonging to the human endogenous retrovirus HERV-W family. We have found that this method may support the PCR technique in screening for minor alleles which, in certain conditions, may be undetected by the standard PCR technique...
January 8, 2018: Molecular Biotechnology
Hélio Nitta Matsuura, Sonia Malik, Fernanda de Costa, Morteza Yousefzadi, Mohammad Hossein Mirjalili, Randolph Arroo, Avninder S Bhambra, Miroslav Strnad, Mercedes Bonfill, Arthur Germano Fett-Neto
Plant secondary metabolism evolved in the context of highly organized and differentiated cells and tissues, featuring massive chemical complexity operating under tight environmental, developmental and genetic control. Biotechnological demand for natural products has been continuously increasing because of their significant value and new applications, mainly as pharmaceuticals. Aseptic production systems of plant secondary metabolites have improved considerably, constituting an attractive tool for increased, stable and large-scale supply of valuable molecules...
December 30, 2017: Molecular Biotechnology
Sudip Biswas, U S Mahzabin Amin, Sarah Sarker, M Sazzadur Rahman, Ruhul Amin, Rezaul Karim, Narendra Tuteja, Zeba I Seraj
DNA helicase (PDH45) from the pea plant (Pisum sativum) is a member of the DEAD box protein family and plays a vital regulatory role in saline stress tolerance in plants. We previously reported that over-expression of PDH45 gene confers both seedling and reproductive stage salinity tolerance to a Bangladeshi rice landrace, Binnatoa (BA). In this study, transgenic BA-containing PDH45 (♂) was crossed with two different farmer-popular BRRI rice varieties (♀), BR28 and BR47, in a contained net house. F1 plants positive for the transgene and having recipient phenotype were advanced from F1 to F5...
December 27, 2017: Molecular Biotechnology
Alireza Ebrahiminezhad, Alireza Zare-Hoseinabadi, Ajit K Sarmah, Saeed Taghizadeh, Younes Ghasemi, Aydin Berenjian
Nanoscale iron particles have attracted substantial interest due to their unique physical and chemical properties. Over the years, various physical and chemical methods have been developed to synthesize these nanostructures which are usually expensive and potentially harmful to human health and the environment. Synthesis of iron nanoparticles (INPs) by using plant extract is now of great interest in order to develop a novel and sustainable approach toward green chemistry. In this method the chemical compounds and organic solvents are replaced with phytochemicals and aqueous matrixes, respectively...
December 18, 2017: Molecular Biotechnology
Maryam Fard, Maryam Akhavan-Tavakoli, Sayeh Khanjani, Sona Zare, Haleh Edalatkhah, Shaghayegh Arasteh, Davood Mehrabani, Amir-Hassan Zarnani, Somaieh Kazemnejad, Reza Shirazi
The skin provides a dynamic barrier separating and protecting human body from the exterior world, and then immediate repair and rebuilding of the epidermal barrier is crucial after wound and injury. Wound healing without scars and complete regeneration of skin tissue still remain as a clinical challenge. The demand to engineer scaffolds that actively promote regeneration of damaged areas of the skin has been increased. In this study, menstrual blood-derived stem cells (MenSCs) have been induced to differentiate into keratinocytes-like cells in the presence of human foreskin-derived keratinocytes on a bilayer scaffold based on amniotic membrane and silk fibroin...
December 15, 2017: Molecular Biotechnology
Yilan Liu, Jinjin Chen, Anders Thygesen
Gene splicing by fusion PCR is a versatile and widely used methodology, especially in synthetic biology. We here describe a rapid method for splicing two fragments by one-round fusion PCR with a dual-asymmetric primers and two-step annealing (ODT) method. During the process, the asymmetric intermediate fragments were generated in the early stage. Thereafter, they were hybridized in the subsequent cycles to serve as template for the target full-length product. The process parameters such as primer ratio, elongation temperature and cycle numbers were optimized...
December 11, 2017: Molecular Biotechnology
Chuannan Long, Yijin Cheng, Jingjing Cui, Jian Liu, Lihui Gan, Bin Zeng, Minnan Long
The role of the transcription factor creA-mediating carbon catabolite repression in Trichoderma orientalis EU7-22 was investigated for cellulase and hemicellulase production. The binary vector pUR5750G/creA::hph was constructed to knock out creA by homologous integration, generating the ΔcreA mutant Trichoderma orientalis CF1D. For strain CF1D, the filter paper activities (FPA), endoglucanase activities (CMC), cellobiohydrolase activity(CBH), β-glucosidase activity (BG), xylanase activity (XYN), and extracellular protein concentration were 1...
December 6, 2017: Molecular Biotechnology
Hirak Ranjan Dash, Surajit Das
Forensic biology is a sub-discipline of biological science with an amalgam of other branches of science used in the criminal justice system. Any nucleated cell/tissue harbouring DNA, either live or dead, can be used as forensic exhibits, a source of investigation through DNA typing. These biological materials of human origin are rich source of proteins, carbohydrates, lipids, trace elements as well as water and, thus, provide a virtuous milieu for the growth of microbes. The obstinate microbial growth augments the degradation process and is amplified with the passage of time and improper storage of the biological materials...
December 6, 2017: Molecular Biotechnology
Saki Yokota, Yurina Suzuki, Keisuke Saitoh, Sakihito Kitajima, Norimasa Ohya, Takeshi Gotoh
Rubber elongation factor (REF) and small rubber particle protein (SRPP) are major latex proteins harvested from Hevea brasiliensis (the rubber tree; HbREF and HbSRPP, respectively). Their amino acid sequences exhibit high homology with each other. In the present study, we cloned two cDNAs encoding REF/SRPP-family proteins (FcREF/SRPP-1 and -2) from the laticifers of Ficus carica (fig tree). The amino acid sequences of these proteins showed high homology not only with each other but also with HbREF and HbSRPP...
December 6, 2017: Molecular Biotechnology
Pouya Dini, Shavahn C Loux, Kirsten E Scoggin, Alejandro Esteller-Vico, Edward L Squires, Mats H T Troedsson, Peter Daels, Barry A Ball
MicroRNAs (miRNAs) have important posttranscriptional regulatory abilities, and there is considerable interest in evaluating their expression patterns in different pathophysiological states. The most common method of quantifying miRNA expression is quantitative reverse transcription PCR; however, the identification of tissue-specific and species-specific reference miRNA is a prerequisite for miRNA expression analysis. Currently, no reference genes have been described for evaluating miRNA expression in equine serum and chorioallantoic membrane (CAM) during pregnancy...
December 2, 2017: Molecular Biotechnology
Przemysław Sitarek, Tomasz Kowalczyk, Patricia Rijo, Adam J Białas, Marzena Wielanek, Halina Wysokińska, Catarina Garcia, Monika Toma, Tomasz Śliwiński, Ewa Skała
This study examines the production of five phenolic acids (chlorogenic acid, neochlorogenic acid, ferulic acid, caffeic acid and p-coumaric acid) following over-expression of AtPAP1 transcription factor by four transgenic root clones of Leonurus sibiricus after Agrobacterium rhizogenes transformation. The AtPAP1 expression level was estimated by quantitative real-time PCR. High levels of phenolic acids were found in the transgenic roots of L. sibiricus and were determined by high-performance liquid chromatography-mass spectrometry analysis...
December 1, 2017: Molecular Biotechnology
Elahe Tavakol
Among the available reverse genetic approaches for studying gene function, virus-induced gene silencing (VIGS) has several advantages. It allows rapid characterization of gene function independent of stable transformation, which is basically difficult to achieve in monocots, and offers the potential to silence individual or multiple genes of a gene family. In order to establish a VIGS system in Aegilops tauschii, modified vectors derived from Barley stripe mosaic virus (BSMV) were used for silencing a phytoene desaturase gene that provides a convenient visual reporter for silencing...
December 1, 2017: Molecular Biotechnology
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