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Molecular Biotechnology

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https://www.readbyqxmd.com/read/29987520/transcriptional-study-of-the-rsmz-srnas-and-their-relationship-to-the-biosynthesis-of-alginate-and-alkylresorcinols-in-azotobacter-vinelandii
#1
Liliana López-Pliego, Liliana García-Ramírez, Emma Aurora Cruz-Gómez, Patricia Domínguez-Ojeda, Alejandra López-Pastrana, Luis Ernesto Fuentes-Ramírez, Cinthia Núñez, Miguel Castañeda
The GacS/A system in Azotobacter vinelandii regulates alginate and alkylresorcinols production through RsmZ1, a small regulatory RNA (sRNA) that releases the translational repression of the algD and arpR mRNAs caused by the RsmA protein. In the Pseudomonadaceae family, the Rsm-sRNAs are grouped into three families: RsmX, RmsY and RsmZ. Besides RsmZ1, A. vinelandii has six other isoforms belonging to the RsmZ family and another one to the RsmY. Environmental signals controlling rsmsRNAs genes in A. vinelandii are unknown...
July 10, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29974328/a-graft-mimic-strategy-for-verticillium-resistance-in-tomato
#2
Melora Mackey, Alexander Kurosky, E Jane Robb, Ross N Nazar
Grafting vegetables for disease resistance has increased greatly in popularity over the past 10 years. Verticillium wilt of tomato is commonly controlled through grafting of commercial varieties on resistant rootstocks expressing the Ve1 R-gene. To mimic the grafted plant, proteomic analyses in tomato were used to identify a suitable root-specific promoter (TMVi), which was used to express the Ve1-allele in susceptible Craigella (Cs) tomato plants. The results indicate that when infected with Verticillim dahliae, race 1, the transformed plants are comparable to resistant cultivars (Cr) or grafted plants...
July 4, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29951737/the-development-of-bacterial-carboxylesterase-biological-recognition-elements-for-cocaine-detection
#3
Suhad A Mustafa
Enzyme recognition element-based biosensors are very attractive for biosensor application due to a variety of measurable reaction products arising from a catalytic process. In this study, biosensor recognition elements have been developed via engineer bacterial enzymes (carboxylesterases (CEs)) which will used for narcotic detection because of their role in narcotics metabolism. The modification (insertion of cys-tag) allows the enzyme to bind into a transducer surface of a biosensor which will translate the reaction product into the detection system...
June 27, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29951736/molecular-cloning-and-biochemical-characterization-of-iron-superoxide-dismutase-from-leishmania-braziliensis
#4
Camila C B Brito, Fernando V Maluf, Gustavo M A de Lima, Rafael V C Guido, Marcelo S Castilho
Leishmaniasis is one of the most important neglected tropical diseases, with a broad spectrum of clinical manifestations. Among the clinical manifestations of the disease, cutaneous leishmaniasis, caused by species of Leishmania braziliensis, presents wide distribution in Brazil. In this work, we performed the cloning, expression, and purification of the enzyme superoxide dismutase of Leishmania braziliensis (LbSOD-B2) considered a promising target for the search of new compounds against leishmaniasis. In vitro assays based on pyrogallol oxidation showed that LbSOD-B2 is most active around pH 8 and hydrogen peroxide is a LbSOD-B2 inhibitor at low millimolar range (IC50  = 1 mM)...
June 27, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29946933/an-update-on-genetic-modification-of-chickpea-for-increased-yield-and-stress-tolerance
#5
REVIEW
Manoj Kumar, Mohd Aslam Yusuf, Manisha Nigam, Manoj Kumar
Chickpea is a highly nutritious grain legume crop, widely appreciated as a health food, especially in the Indian subcontinent. The major constraints on chickpea production are biotic (Helicoverpa, bruchid, aphid, ascochyta) and abiotic (drought, heat, salt, cold) stresses, which reduce the yield by up to 90%. Various strategies like conventional breeding, molecular breeding, and modern plant breeding have been used to overcome these problems. Conventionally, breeding programs aim at development of varieties that combine maximum number of traits through inter-specific hybridization, wide hybridization, and hybridization involving more than two parents...
June 26, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29943150/soluble-production-of-human-recombinant-vegf-a121-by-using-sumo-fusion-technology-in-escherichia-coli
#6
Rufus Vinod Munawar Samuel, Syeda Yumna Farrukh, Sadia Rehmat, Muhammad Umair Hanif, Syed Shoaib Ahmed, Syed Ghulam Musharraf, Faiza Gul Durrani, Mahjabeen Saleem, Roquyya Gul
Human recombinant vascular endothelial growth factor-A121 (hrVEGF-A121) has applications in pharmaceutical industry especially in regenerative medicine. Here, we report the expression, purification, and characterization of hrVEGF-A121 in Escherichia coli expression system using human small ubiquitin-related modifier-3 (hSUMO3) fusion partner. Total RNA was isolated from healthy human gingival tissue, VEGF-A121 gene was RT-PCR amplified, and hSUMO3 gene was tagged at N-terminus. The fusion gene (SUMO3-VEGF-A121) was cloned in pET-22b(+) expression vector and transferred into E...
June 25, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29943149/combinatorial-interactions-of-biotic-and-abiotic-stresses-in-plants-and-their-molecular-mechanisms-systems-biology-approach
#7
REVIEW
Arun Kumar Dangi, Babita Sharma, Ishu Khangwal, Pratyoosh Shukla
Plants are continually facing biotic and abiotic stresses, and hence, they need to respond and adapt to survive. Plant response during multiple and combined biotic and abiotic stresses is highly complex and varied than the individual stress. These stresses resulted alteration of plant behavior through regulating the levels of microRNA, heat shock proteins, epigenetic variations. These variations can cause many adverse effects on the growth and development of the plant. Further, in natural conditions, several abiotic stresses causing factors make the plant more susceptible to pathogens infections and vice-versa...
June 25, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29943148/engineering-yarrowia-lipolytica-for-use-in-biotechnological-applications-a-review-of-major-achievements-and-recent-innovations
#8
REVIEW
Catherine Madzak
Yarrowia lipolytica is an oleaginous saccharomycetous yeast with a long history of industrial use. It aroused interest several decades ago as host for heterologous protein production. Thanks to the development of numerous molecular and genetic tools, Y. lipolytica is now a recognized system for expressing heterologous genes and secreting the corresponding proteins of interest. As genomic and transcriptomic tools increased our basic knowledge on this yeast, we can now envision engineering its metabolic pathways for use as whole-cell factory in various bioconversion processes...
June 25, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29943147/effect-of-temporal-expression-of-integral-membrane-proteins-by-baculovirus-expression-vector-system
#9
T Z Salem, F Zhang, N Sahly, S Thiem
Integral membrane proteins (IMPs) are popular target for drugs, but their resolved structures have been overlooked when compared with cytosolic proteins. The main reason is that IMPs usually need intensive post-translational modifications and they are bound to membranes, which increase the complexity of purifying or crystalizing them. Although different expression systems are used to express IMPs, baculovirus is considered one of the most successful expression systems for those proteins. Despite that, there are always unknown discrepancies in the level of IMPs expression in the baculovirus expression system...
June 25, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29936696/trichomonas-vaginalis-metalloproteinase-tvmp50-is-a-monomeric-aminopeptidase-p-like-enzyme
#10
Rodrigo Arreola, José Luis Villalpando, Jonathan Puente-Rivera, Jorge Morales-Montor, Enrique Rudiño-Piñera, María Elizbeth Alvarez-Sánchez
Previously, metalloproteinase was isolated and identified from Trichomonas vaginalis, belonging to the aminopeptidase P-like metalloproteinase subfamily A/B, family M24 of clan MG, named TvMP50. The native and recombinant TvMP50 showed proteolytic activity, determined by gelatin zymogram, and a 50 kDa band, suggesting that TvMP50 is a monomeric active enzyme. This was an unexpected finding since other Xaa-Pro aminopeptidases/prolidases are active as a biological unit formed by dimers/tetramers. In this study, the evolutionary history of TvMP50 and the preliminary crystal structure of the recombinant enzyme determined at 3...
June 23, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29916131/generation-of-gtko-diannan-miniature-pig-expressing-human-complementary-regulator-proteins-hcd55-and-hcd59-via-t2a-peptide-based-bicistronic-vectors-and-scnt
#11
Fengjuan Liu, Jinji Liu, Zaimei Yuan, Yubo Qing, Honghui Li, Kaixiang Xu, Wanyun Zhu, Heng Zhao, Baoyu Jia, Weirong Pan, Jianxiong Guo, Xuezeng Zhang, Wenmin Cheng, Wei Wang, Hong-Ye Zhao, Hong-Jiang Wei
Pig-to-human organ transplantation has drawn attention in recent years due to the potential use of pigs as an alternative source of human donor organs. While GGTA1 knockout (GTKO) can protect xenografts from hyperacute rejection, complement-dependent cytotoxicity might still contribute to this type of rejection. To prolong the xenograft survival, we utilized a T2A-mediated pCMV-hCD55-T2A-hCD59-Neo vector and transfected the plasmid into GTKO Diannan miniature pig fetal fibroblasts. After G418 selection combined with single-cell cloning culture, four colonies were obtained, and three of these were successfully transfected with the hCD55 and hCD59...
June 18, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29876740/application-of-nanobit-for-monitoring-dimerization-of-the-null-hong-kong-variant-of-%C3%AE-1-antitrypsin-nhk-in-living-cells
#12
Junpei Norisada, Keito Fujimura, Fumimasa Amaya, Hiroki Kohno, Yoko Hirata, Kentaro Oh-Hashi
In this study, we investigated expression and dimerization of an ER-associated degradation (ERAD) substrate, a null Hong Kong variant of α-1-antitrypsin (NHK) using immunoblotting assay and a novel NanoLuc complementary reporter system called the NanoBiT (NB) assay. This NB-tagged NHK made it possible to monitor the intra- and extracellular status of NHK in living cells. The values for this NB assay fluctuated in response to distinct pharmacological stimuli and co-transfection of several ERAD-related factors...
June 6, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29860626/pellino-1-protects-periodontal-ligament-stem-cells-against-h-2-o-2-induced-apoptosis-via-activation-of-nf-%C3%AE%C2%BAb-signaling
#13
Jiangang Tian, Liufang Gu, Andrew Adams, Xueliang Wang, Ruizhe Huang
To determine the protective effects of Pellino-1 against H2 O2 -induced apoptosis in periodontal ligament stem cells (PDLSC). We demonstrated that H2 O2 decreases PDLSC viability by 40 and 50% with the concentrations of 400 and 500 μM, respectively, with an observed downregulation of Pellino-1 mRNA and protein; we further concluded that overexpression of Pellino-1 significantly lowers 8-hydroxy-2'-deoxyguanosine levels by 10% and upregulates superoxide dismutase 1, glutathione peroxidase levels, and catalase mRNA levels by 200, 40, and 250%, respectively...
June 2, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29855997/synthetic-promoters-designing-the-cis-regulatory-modules-for-controlled-gene-expression
#14
REVIEW
Jameel Aysha, Muhammad Noman, Fawei Wang, Weican Liu, Yonggang Zhou, Haiyan Li, Xiaowei Li
Designing the expression cassettes with desired properties remains the most important consideration of gene engineering technology. One of the challenges for predictive gene expression is the modeling of synthetic gene switches to regulate one or more target genes which would directly respond to specific chemical, environmental, and physiological stimuli. Assessment of natural promoter, high-throughput sequencing, and modern biotech inventory aided in deciphering the structure of cis elements and molding the native cis elements into desired synthetic promoter...
May 31, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29761314/pcl-based-composite-scaffold-matrices-for-tissue-engineering-applications
#15
REVIEW
Nadeem Siddiqui, Simran Asawa, Bhaskar Birru, Ramaraju Baadhe, Sreenivasa Rao
Biomaterial-based scaffolds are important cues in tissue engineering (TE) applications. Recent advances in TE have led to the development of suitable scaffold architecture for various tissue defects. In this narrative review on polycaprolactone (PCL), we have discussed in detail about the synthesis of PCL, various properties and most recent advances of using PCL and PCL blended with either natural or synthetic polymers and ceramic materials for TE applications. Further, various forms of PCL scaffolds such as porous, films and fibrous have been discussed along with the stem cells and their sources employed in various tissue repair strategies...
May 14, 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29796788/bio-production-of-baccatin-iii-an-important-precursor-of-paclitaxel-by-a-cost-effective-approach
#16
Shu-Ling Lin, Tao Wei, Jun-Fang Lin, Li-Qiong Guo, Guang-Pei Wu, Jun-Bin Wei, Jia-Jun Huang, Ping-Lan Ouyang
Natural production of anti-cancer drug taxol from Taxus has proved to be environmentally unsustainable and economically unfeasible. Currently, bioengineering the biosynthetic pathway of taxol is an attractive alternative production approach. 10-deacetylbaccatin III-10-O-acetyl transferase (DBAT) was previously characterized as an acyltransferase, using 10-deacetylbaccatin III (10-DAB) and acetyl CoA as natural substrates, to form baccatin III in the taxol biosynthesis. Here, we report that other than the natural acetyl CoA (Ac-CoA) substrate, DBAT can also utilize vinyl acetate (VA), which is commercially available at very low cost, acylate quickly and irreversibly, as acetyl donor in the acyl transfer reaction to produce baccatin III...
July 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29785699/spectroscopic-analysis-of-the-cu-2-induced-fluorescence-quenching-of-fluorescent-proteins-amcyan-and-morange2
#17
Ji-Eun Bae, In Jung Kim, Ki Hyun Nam
Fluorescent proteins show fluorescence quenching by specific metal ions, which can be applied towards metal biosensing applications. In order to develop metal-biosensor, we performed spectroscopic analysis of the fluorescence quenching of fluorescent protein AmCyan and mOrange2 by various metal ions. The fluorescence intensity of AmCyan was reduced to 48.54% by Co2+ and 67.77% by Zn2+ ; Cu2+ reduced the fluorescence emission of AmCyan to 19.30% of its maximum. The fluorescence intensity of mOrange2 was quenched by only Cu2+ , to 11...
July 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29779127/physiological-and-transcriptomic-analysis-of-a-chronologically-long-lived-saccharomyces-cerevisiae-strain-obtained-by-evolutionary-engineering
#18
Mevlüt Arslan, Can Holyavkin, Halil İbrahim Kısakesen, Alican Topaloğlu, Yusuf Sürmeli, Zeynep Petek Çakar
High-throughput aging studies with yeast as a model organism involve transposon-mutagenesis and yeast knockout collection, which have been pivotal strategies for understanding the complex cellular aging process. In this study, a chronologically long-lived Saccharomyces cerevisiae mutant was successfully obtained by using another high-throughput approach, evolutionary engineering, based on systematic selection in successive batch cultures under gradually increasing levels of caloric restriction. Detailed comparative physiological and transcriptomic analyses of the chronologically long-lived mutant and the reference strain revealed enhanced levels of respiratory metabolism, upregulation of genes related to carbohydrate metabolic processes, glycogen-trehalose pathways, stress response, and repression of protein synthesis-related genes in the long-lived mutant SRM11, already in the absence of caloric restriction...
July 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29730712/homing-genes-expression-in-fucosyltransferase-vi-treated-umbilical-cord-blood-cd133-cells-which-expanded-on-protein-coated-nanoscaffolds
#19
Amir Atashi, Maryam Islami, Yousef Mortazavi, Masoud Soleimani
Umbilical cord blood (UCB)-derived hematopoietic stem cells (HSCs) are considered because of their self-renewing, differentiating, proliferating, and readily available properties. Moreover, HSCs' homing to the hematopoietic microenvironment is an important step in their transplantation process. But low content of progenitor cells in one unit of UCB and defect in the bone marrow (BM) homing limit their applications. Hence, we decided to correct this deficiency with ex vivo incubation of CD133+ cells using fucosyltransferase VI and GDP-fucose...
July 2018: Molecular Biotechnology
https://www.readbyqxmd.com/read/29704159/diversity-of-immunoglobulin-ig-isotypes-and-the-role-of-activation-induced-cytidine-deaminase-aid-in-fish
#20
REVIEW
Bhakti Patel, Rajanya Banerjee, Mrinal Samanta, Surajit Das
The disparate diversity in immunoglobulin (Ig) repertoire has been a subject of fascination since the emergence of prototypic adaptive immune system in vertebrates. The carboxy terminus region of activation-induced cytidine deaminase (AID) has been well established in tetrapod lineage and is crucial for its function in class switch recombination (CSR) event of Ig diversification. The absence of CSR in the paraphyletic group of fish is probably due to changes in catalytic domain of AID and lack of cis-elements in IgH locus...
June 2018: Molecular Biotechnology
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