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Molecular Biotechnology

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https://www.readbyqxmd.com/read/28721592/construction-purification-and-characterization-of-a-homodimeric-granulocyte-colony-stimulating-factor
#1
Gitana Mickiene, Indre Dalgediene, Zilvinas Dapkunas, Gintautas Zvirblis, Henrikas Pesliakas, Algirdas Kaupinis, Mindaugas Valius, Edita Mistiniene, Milda Pleckaityte
Granulocyte colony-stimulating factor (G-CSF) has found widespread clinical application, and modified forms with improved biopharmaceutical properties have been marketed as well. PEGylation, the covalent modification of G-CSF with polyethylene glycol (PEG), has a beneficial effect on drug properties, but there are concerns connected to the immunogenicity of PEGylated compounds and bioaccumulation of the synthetic polymer. To overcome challenges connected with chemical modifications, we developed fusion proteins composed of two G-CSF molecules connected via different peptide linkers...
July 18, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28698981/genetically-modified-pigs-as-organ-donors-for-xenotransplantation
#2
REVIEW
Magdalena Hryhorowicz, Joanna Zeyland, Ryszard Słomski, Daniel Lipiński
The growing shortage of available organs is a major problem in transplantology. Thus, new and alternative sources of organs need to be found. One promising solution could be xenotransplantation, i.e., the use of animal cells, tissues and organs. The domestic pig is the optimum donor for such transplants. However, xenogeneic transplantation from pigs to humans involves high immune incompatibility and a complex rejection process. The rapid development of genetic engineering techniques enables genome modifications in pigs that reduce the cross-species immune barrier...
July 11, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28695481/a-novel-rat-model-of-nonalcoholic-fatty-liver-disease-constructed-through-crispr-cas-based-hydrodynamic-injection
#3
Qin Yu, Rui-Zhi Tan, Quan Gan, Xia Zhong, You-Qiang Wang, Jing Zhou, Li Wang
CRISPR/Cas technology has been widely used in generating conventional and conditional gene knockout animals through germline mutation. A recent study has demonstrated that CRISPR/Cas system also worked on nongermline mutation in mice liver via hydrodynamic injection of vector plasmid to blood circulation. However, whether this is also applied to rat and what is the optimal concentration of vector plasmid still need to be studied. Here, we attempted to use pX330-Pten plasmid to knock down the expression of Pten in rat liver for generation of nonalcoholic fatty liver disease (NAFLD) rat model...
July 10, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28688042/erratum-to-species-specific-detection-of-mycosphaerella-polygoni-cuspidati-as-a-biological-control-agent-for-fallopia-japonica-by-pcr-assay
#4
Daisuke Kurose, Naruto Furuya, Tetsuya Saeki, Kenichi Tsuchiya, Seiya Tsushima, Marion K Seier
No abstract text is available yet for this article.
July 7, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28676913/co-expression-of-beta-glucosidase-and-laccase-in-trichoderma-reesei-by-random-insertion-with-enhanced-filter-paper-activity
#5
Ruowen Wang, Jing Yang, Guoqing Zhang, Yapeng Chao, Zhimin Li, Qin Ye, Shijun Qian
Trichoderma reesei strain Rut-C30 was modified with enhanced beta-glycosidase (BGL) activity to balance the cellulase system and generated laccase (LAC) protein for lignin degradation. Initially, the binary plasmid p1300-w1 was constructed to express T. reesei bgl2 under the control of promoter P pki and T-nos terminator. Random insertion was performed via Agrobacterium tumefaciens-mediated transformation. A total of 353 mutants were obtained, and 34PTrb2 was exceptionally stable with increased FPA and BGL activity after screening for extracellular enzyme activity...
July 4, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28674943/host-induced-silencing-of-pathogenicity-genes-enhances-resistance-to-fusarium-oxysporum-wilt-in-tomato
#6
Poonam Bharti, Poonam Jyoti, Priya Kapoor, Vandana Sharma, V Shanmugam, Sudesh Kumar Yadav
This study presents a novel approach of controlling vascular wilt in tomato by RNAi expression directed to pathogenicity genes of Fusarium oxysporum f. sp. lycopersici. Vascular wilt of tomato caused by Fusarium oxysporum f. sp. lycopersici leads to qualitative and quantitative loss of the crop. Limitation in the existing control measures necessitates the development of alternative strategies to increase resistance in the plants against pathogens. Recent findings paved way to RNAi, as a promising method for silencing of pathogenicity genes in fungus and provided effective resistance against fungal pathogens...
July 3, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28667571/extracellular-secretion-of-phytase-from-transgenic-wheat-roots-allows-utilization-of-phytate-for-enhanced-phosphorus-uptake
#7
Samreen Mohsin, Asma Maqbool, Mehwish Ashraf, Kauser Abdulla Malik
A significant portion of organic phosphorus comprises of phytates which are not available to wheat for uptake. Hence for enabling wheat to utilize organic phosphorus in form of phytate, transgenic wheat expressing phytase from Aspergillus japonicus under barley root-specific promoter was developed. Transgenic events were initially screened via selection media containing BASTA, followed by PCR and BASTA leaf paint assay after hardening. Out of 138 successfully regenerated To events, only 12 had complete constructs and thus further analyzed...
June 30, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28667570/molecular-cloning-expression-and-characterization-of-oenococcus-oeni-priming-glycosyltransferases
#8
Maria Dimopoulou, Olivier Claisse, Lucie Dutilh, Cécile Miot-Sertier, Patricia Ballestra, Patrick M Lucas, Marguerite Dols-Lafargue
Oenococcus oeni is the main bacterial species that drives malolactic fermentation in wine. Most O. oeni strains produce capsular exopolysaccharides (EPS) that may contribute to protect them in the wine hostile environment. In O. oeni genome sequences, several genes are predicted to encode priming glycosyltransferases (pGTs). These enzymes are essential for EPS formation as they catalyze the first biosynthetic step through the formation of a phosphoanhydride bond between a hexose-1-phosphate and a lipid carrier undecaprenyl phosphate...
June 30, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28664508/correlation-between-expression-of-recombinant-proteins-and-abundance-of-h3k4me3-on-the-enhancer-of-human-cytomegalovirus-major-immediate-early-promoter
#9
Benjamin P C Soo, Julian Tay, Shirelle Ng, Steven C L Ho, Yuansheng Yang, Sheng-Hao Chao
Role of epigenetic regulation in the control of gene expression is well established. The impact of several epigenetic mechanisms, such as DNA methylation and histone acetylation, on recombinant protein production in mammalian cells has been investigated recently. Here we investigate the correlation between the selected epigenetic markers and five trastuzumab biosimilar-producing Chinese hamster ovary (CHO) cell lines in which the expression of trastuzumab is driven by human cytomegalovirus (HCMV) major immediate-early (MIE) promoter...
June 29, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28631058/expression-and-characterization-of-glucose-oxidase-from-aspergillus-niger-in-yarrowia-lipolytica
#10
Fatemeh Khadivi Derakshan, Farshad Darvishi, Mehrouz Dezfulian, Catherine Madzak
Glucose oxidase (GOX) is currently used in clinical, pharmaceutical, food and chemical industries. The aim of this study was expression and characterization of Aspergillus niger glucose oxidase gene in the yeast Yarrowia lipolytica. For the first time, the GOX gene of A. niger was successfully expressed in Y. lipolytica using a mono-integrative vector containing strong hybrid promoter and secretion signal. The highest total glucose oxidase activity was 370 U/L after 7 days of cultivation. An innovative method was used to cell wall disruption in current study, and it could be recommended to use for efficiently cell wall disruption of Y...
June 19, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28597202/erratum-to-design-of-a-lentiviral-vector-for-the-inducible-expression-of-myc-a-new-strategy-for-construction-approach
#11
Onur Tokgun, Francesco Paolo Fiorentino, Pervin Elvan Tokgun, Jun Yokota, Hakan Akca
No abstract text is available yet for this article.
June 8, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28585118/analysis-of-promoters-of-arabidopsis-thaliana-divergent-gene-pair-serat3-2-and-idh-iii-shows-serat3-2-promoter-is-nested-within-the-idh-iii-promoter
#12
Ritesh Kumar Raipuria, Vajinder Kumar, Kadur Narayan Guruprasad, Shripad Ramachandra Bhat
Intergenic regions of divergent gene pairs show bidirectional promoter activity but whether regulatory sequences for gene expression in opposite directions are shared is not established. In this study, promoters of divergently arranged gene pair At4g35640-At4g35650 (SERAT3;2-IDH-III) of Arabidopsis thaliana were analyzed to identify overlapping regulatory regions. Both genes showed the highest expression in flower buds and flowers. 5' RACE experiments extended the intergenic region from 161 bp shown in TAIR annotation to 512 bp...
June 5, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28580552/ability-of-t1-lipase-to-degrade-amorphous-p-3hb-structural-and-functional-study
#13
Rauda A Mohamed, Abu Bakar Salleh, Adam Thean Chor Leow, Normi M Yahaya, Mohd Basyaruddin Abdul Rahman
An enzyme with broad substrate specificity would be an asset for industrial application. T1 lipase apparently has the same active site residues as polyhydroxyalkanoates (PHA) depolymerase. Sequences of both enzymes were studied and compared, and a conserved lipase box pentapeptide region around the nucleophilic serine was detected. The alignment of 3-D structures for both enzymes showed their active site residues were well aligned with an RMSD value of 1.981 Å despite their sequence similarity of only 53...
June 3, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28573450/cloning-production-and-characterization-of-a-glycoside-hydrolase-family-7-enzyme-from-the-gut-microbiota-of-the-termite-coptotermes-curvignathus
#14
James Sy-Keen Woon, Patricia Jie Hung King, Mukram Mohamed Mackeen, Nor Muhammad Mahadi, Wan Mohd Khairulikhsan Wan Seman, William J Broughton, Abdul Munir Abdul Murad, Farah Diba Abu Bakar
Coptotermes curvignathus is a termite that, owing to its ability to digest living trees, serves as a gold mine for robust industrial enzymes. This unique characteristic reflects the presence of very efficient hydrolytic enzyme systems including cellulases. Transcriptomic analyses of the gut of C. curvignathus revealed that carbohydrate-active enzymes (CAZy) were encoded by 3254 transcripts and that included 69 transcripts encoding glycoside hydrolase family 7 (GHF7) enzymes. Since GHF7 enzymes are useful to the biomass conversion industry, a gene encoding for a GHF7 enzyme (Gh1254) was synthesized, sub-cloned and expressed in the methylotrophic yeast Pichia pastoris...
June 1, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28573449/medium-optimization-for-improved-production-of-dihydrolipohyl-dehydrogenase-from-bacillus-sphaericus-pad-91-in-escherichia-coli
#15
Hamid Shahbazmohammadi, Eskandar Omidinia
Dihydrolipohyl dehydrogenase (DLD) is a FAD-dependent enzyme that catalyzes the reversible oxidation of dihydrolipoamide. Herein, we report medium optimization for the production of a recombinant DLD with NADH-dependent diaphorase activity from a strain of Bacillus sphaericus PAD-91. The DLD gene that consisted of 1413 bp was expressed in Escherichia coli BL21 (DE3), and its enzymatic properties were studied. The composition of production medium was optimized using one-variable-at-a-time method followed by response surface methodology (RSM)...
June 1, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28567686/tobacco-rattle-virus-based-silencing-of-enoyl-coa-reductase-gene-and-its-role-in-resistance-against-cotton-wilt-disease
#16
Roma Mustafa, Muhammad Hamza, Hira Kamal, Shahid Mansoor, Jodi Scheffler, Imran Amin
A Tobacco rattle virus (TRV)-based virus-induced gene silencing assay was employed as a reverse genetic approach to study gene function in cotton (Gossypium hirsutum). This approach was used to investigate the function of the Enoyl-CoA reductase (GhECR) gene in pathogen defense. Amino acid sequence alignment of Arabidopsis ECR with homologous sequence from G. hirsutum, G. arboreum, G. herbaceum and G. barbadense showed that ECRs are highly conserved among these species. TRV-based silencing of GhECR gene in G...
May 31, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28526937/molecular-confirmation-of-intraspecific-tomato-solanum-lycopersicum-hybrids-and-their-evaluation-against-late-blight-and-cucumber-mosaic-virus
#17
Amjad Hameed, Muhammad Yussouf Saleem, Khalid Pervaiz Akhtar, Muhammad Shoaib, Qumer Iqbal, Muhammad Asghar
Tomato is one of the most consumed vegetables in the world. Diseases are the number one concern in the development of high-yield and disease-resistant tomato hybrids which is the foremost priority of breeders. Present study was conducted (1) to develop DNA-based markers for genetic confirmation of tomato F1 hybrids, (2) to utilize sequenced characterized amplified region (SCAR) marker linked to the Ph-3 gene for Phytophthora infestans resistance in tomato and (3) to evaluate male and female parental genotypes and their F1 hybrids against late blight (LB) and cucumber mosaic virus (CMV)...
May 19, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28509990/transient-activation-of-reprogramming-transcription-factors-using-protein-transduction-facilitates-conversion-of-human-fibroblasts-toward-cardiomyocyte-like-cells
#18
Zaniar Ghazizadeh, Hassan Rassouli, Hananeh Fonoudi, Mehdi Alikhani, Mahmood Talkhabi, Amir Darbandi-Azar, Shuibing Chen, Hossein Baharvand, Nasser Aghdami, Ghasem Hosseini Salekdeh
Derivation of cardiomyocytes directly from patients' own fibroblasts could offer a new therapeutic approach for those with ischemic heart disease. An essential step toward clinical application is to establish safe conversion of human fibroblasts into a cardiac fate. Here we aimed to efficiently and safely generate cardiomyocytes from human fibroblasts by direct delivery of reprogramming recombinant cell permeant form of reprogramming proteins followed by cardio-inductive signals. Human fetal and adult fibroblasts were transiently exposed to transactivator of transcription-fused recombinant OCT4, SOX2, KLF4 and c-MYC for 2 weeks and then were directly differentiated toward protein-induced cardiomyocyte-like cells (p-iCLCs) in a cardiac fate niche, carried out by treatment with a set of cardiogenic small molecules (sequential treatment of Chir, and IWP-2, SB431542 and purmorphamine)...
May 16, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28500482/improved-yield-of-high-molecular-weight-hyaluronic-acid-production-in-a-stable-strain-of-streptococcus-zooepidemicus-via-the-elimination-of-the-hyaluronidase-encoding-gene
#19
Navid Pourzardosht, Mohammad Javad Rasaee
Despite the significant potential of Streptococcus zooepidemicus for hyaluronic acid (HA) production with high molecular weight (MW), the HA degrading properties of hyaluronidase prevents the bacteria to achieve enhanced HA yield with high MW. In the present study, we aim to knockout the hyaluronidase enzyme and assess its effects on the yield and MW of the produced HA. The kanamycin resistance gene between the left and right arms of hyaluronidase gene was inserted into pUC18 plasmid to construct pUC18-L-kana(r)-R as a recombinant suicide plasmid...
May 12, 2017: Molecular Biotechnology
https://www.readbyqxmd.com/read/28484957/characterization-of-recombinant-thermococcus-kodakaraensis-kod-dna-polymerases-produced-using-silkworm-baculovirus-expression-vector-system
#20
Mami Yamashita, Jian Xu, Daisuke Morokuma, Kazuma Hirata, Masato Hino, Hiroaki Mon, Masateru Takahashi, Samir M Hamdan, Kosuke Sakashita, Kazuhiro Iiyama, Yutaka Banno, Takahiro Kusakabe, Jae Man Lee
The KOD DNA polymerase from Thermococcus kodakarensis (Tkod-Pol) has been preferred for PCR due to its rapid elongation rate, extreme thermostability and outstanding fidelity. Here in this study, we utilized silkworm-baculovirus expression vector system (silkworm-BEVS) to express the recombinant Tkod-Pol (rKOD) with N-terminal (rKOD-N) or C-terminal (rKOD-C) tandem fusion tags. By using BEVS, we produced functional rKODs with satisfactory yields, about 1.1 mg/larva for rKOD-N and 0.25 mg/larva for rKOD-C, respectively...
May 8, 2017: Molecular Biotechnology
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