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Molecular Biotechnology

Ruowei Li, Guangbo Kang, Min Hu, He Huang
Ribosome display is a powerful engineering research tool for the high-throughput selection of peptides or proteins, which results in the generation of high-performance binders against nearly any antigen of interest. As a cell-free display system, ribosome display has been well developed with many outstanding achievements for over 20 years. Compared with other related display techniques, ribosome display shows unique advantages and development prospects. This tool has been successfully exploited for the selection of functional and specific binders in vitro...
November 7, 2018: Molecular Biotechnology
Asmita Kamble, Sumana Srinivasan, Harinder Singh
Enzymes are essential biological macromolecules, which catalyse chemical reactions and have impacted the human civilization tremendously. The importance of enzymes as biocatalyst was realized more than a century ago by eminent scientists like Kuhne, Buchner, Payen, Sumner, and the last three decades has seen exponential growth in enzyme industry, mainly due to the revolution in tools and techniques in molecular biology, biochemistry and production. This has resulted in high demand of enzymes in various applications like food, agriculture, chemicals, pharmaceuticals, cosmetics, environment and research sector...
November 7, 2018: Molecular Biotechnology
Ali Fathi, Hassan Rasouli, Meghdad Yeganeh, Ghassem Hosseini Salekdeh, Hossein Baharvand
The original version of this article was published without article note. The article note is given below.
November 3, 2018: Molecular Biotechnology
Susan Westfall, Nikita Lomis, Satya Prakash
The composition and activity of the gut microbiota impacts several energy-regulating conditions including diabetes, obesity and metabolic syndrome; however, the specific mechanisms linking the gut microbiota with the host's energy homeostasis remain elusive. Probiotics are health-promoting bacteria that when consumed, alter the composition and/or metabolism of resident microbiota conferring health benefits. To assess the role of a specific probiotic treatment on microbiota-derived impacts on energy homeostasis in the context of development, Drosophila melanogaster larvae were orally administered the probiotic Lactobacillus fermentum NCIMB 5221 or its metabolic product, ferulic acid: a potent anti-inflammatory and anti-oxidant hydroxycinnamic acid...
October 27, 2018: Molecular Biotechnology
Yan-Chao Yin, Xiao-Dong Zhang, Zhi-Qiang Gao, Ting Hu, Ying Liu
Chalcone isomerase (CHI) is the second rate-limiting and the first reported enzyme involved in the biosynthetic pathway of flavonoids. It catalyzes the intramolecular cyclization reaction, converting the bicyclic chalcone into tricyclic (2S)-flavanone. In this paper, we obtained and analyzed 916 DNA sequences, 1310 mRNA sequences, and 2403 amino acid sequences of CHI registered in NCBI by Jan 2018. The full length of CHI DNA sequences ranges from 218 to 3758 bp, CHI mRNA sequences ranges from 265 to 1436 bp, and CHI amino acid sequences ranges from 35 to 465 amino acid residues...
October 15, 2018: Molecular Biotechnology
Riaz Ahmad, Muhammad Akbar Anjum, Waqas Malik
Mango is also known as 'King of Fruits' and is one of the most popular fruit crops in tropical and sub-tropical regions. Characterization through different markers is an important need for mango germplasm conservation and its utilization for various breeding purposes. Description of existing germplasm is essential to detect anticipated traits or genes. Therefore, different markers are applied. Morphological markers comprise those essential traits which can be recorded through naked eye observation and certainly expressed under different agro-climatic conditions...
October 12, 2018: Molecular Biotechnology
Rachel M Woodfint, Erin Hamlin, Kichoon Lee
Animal bioreactors are genetically modified animal systems that have the potential to reduce production cost, and improve production efficiency, of pharmaceutically relevant recombinant proteins. Several species including goats, cattle, rabbits, and avians have been genetically modified to secrete target proteins into milk, egg whites, blood, or other bodily fluids. There are several advantages associated with the use of avians as bioreactor systems. Avians have a short generation time, leading to the quick establishment of a transgenic line and high egg production...
December 2018: Molecular Biotechnology
Yoshiki Morifuji, Jian Xu, Noriko Karasaki, Kazuhiro Iiyama, Daisuke Morokuma, Masato Hino, Akitsu Masuda, Takumi Yano, Hiroaki Mon, Takahiro Kusakabe, Jae Man Lee
Human α1 -antitrypsin (AAT) is the most abundant serine proteinase inhibitor (serpin) in the human plasma. Commercially available AAT for the medications of deficiency of α1 -antitrypsin is mainly purified from human plasma. There is a high demand for a stable and low-cost supply of recombinant AAT (rAAT). In this study, the baculovirus expression vector system using silkworm larvae as host was employed and a large amount of highly active AAT was recovered from the silkworm serum (~ 15 mg/10 ml) with high purity...
December 2018: Molecular Biotechnology
Mikiko Nakamura, Junya Aihara, Hisashi Hoshida, Rinji Akada
Escherichia coli has been used for recombinant protein production for many years. However, no native E. coli promoters have been found for constitutive expression in LB medium. To obtain high-expression E. coli promoters active in LB medium, we inserted various promoter regions upstream of eEmRFP that encodes a red fluorescent protein. Among the selected promoters, only colonies of srlA promoter transformants turned red on LB plate. srlA is a gene that regulates sorbitol utilization. The addition of sorbitol enhanced eEmRFP expression but glucose and other sugars repressed, indicating that srlAp is a sorbitol-enhanced glucose-repressed promoter...
December 2018: Molecular Biotechnology
Pengjie Wu, Huimin Yu, Jin Xu, Jiangli Wu, Awraris Getachew, Yangyang Tu, Zhanbao Guo, Hongyan Jin, Shufa Xu
The aim of this study was to purify the Chinese Sacbrood Virus Beijing Miyun (BJMY-CSBV) from infected Apis cerana larvae, clone structural protein gene VP1 (named BJMY-CSBV-VP1), and investigate its biological information. The result indicated that the capsid of CSBV is of spherical shape. Gene clone experiment showed that the BJMY-CSBV-VP1 gene sequence comprised 945 bp, encoding 315 amino acids with relative molecular weight of 35.59 kDa and isoelectric point 9.38 pI. Phylogenetic analysis of amino acid sequences showed that the BJMY-CSBV-VP1 and LNDD_2015 were grouped together...
December 2018: Molecular Biotechnology
Angelica Jimenez-Rosales, Miriam V Flores-Merino
Proteins are key biomolecules for most biological processes, their function is related to their conformation that is also dictated by their sequence of amino acids. Through evolution, nature has produced an immense variety of enzymatic tools of high efficiency and selectivity, and thanks to the understanding of the molecular basis of life and the technological advances, scientists have learned to introduce mutations and select mutant enzymes, to optimize and control their molecular fitness characteristics mainly for industrial, medical and environmental applications...
December 2018: Molecular Biotechnology
Liliana Morales, Paula Hernández, Jacqueline Chaparro-Olaya
Constructs containing partial coding sequences of myosin A, myosin B, and glideosome-associated protein (50 kDa) of Plasmodium falciparum were used to challenge several strategies designed in order to improve the production and solubility of recombinant proteins in Escherichia coli. Assays were carried out inducing expression in a late log phase culture, optimizing the inductor concentration, reducing the growth temperature for induced cultures, and supplementing additives in the lysis buffer. In addition, recombinant proteins were expressed as fusion proteins with three different tags (6His, GST, and MBP) in four different E...
December 2018: Molecular Biotechnology
Wei Tang, Caroline Luo
RNA editing is a fundamental biochemical process relating to the modification of nucleotides in messenger RNAs of functional genes in cells. RNA editing leads to re-establishment of conserved amino acid residues for functional proteins in nuclei, chloroplasts, and mitochondria. Identification of RNA editing factors that contributes to target site recognition increases our understanding of RNA editing mechanisms. Significant progress has been made in recent years in RNA editing studies for both animal and plant cells...
December 2018: Molecular Biotechnology
Natalia A Kolganova, Vadim A Vasiliskov, Viktoriya E Kuznetsova, Valeriy E Shershov, Sergey A Lapa, Timur O Guseinov, Maksim A Spitsyn, Edward N Timofeev, Alexander V Chudinov
The transferase activity of non-proofreading DNA polymerases is a well-known phenomenon that has been utilized in cloning and sequencing applications. The non-templated addition of modified nucleotides at DNA blunt ends is a potentially useful feature of DNA polymerases that can be used for selective transformation of DNA 3' ends. In this paper, we characterized the tailing reaction at perfectly matched and mismatched duplex ends with Cy3- and Cy5-modified pyrimidine nucleotides. It was shown that the best DNA tailing substrate does not have a perfect Watson-Crick base pair at the end...
December 2018: Molecular Biotechnology
Arezoo Golestanipour, Maryam Nikkhah, Ali Aalami, Saman Hosseinkhani
Development of efficient, easy, and safe gene delivery methods is of great interest in the field of plant biotechnology. Considering the limitations of the usual transfection methods (such as transgene size and plant type), several new techniques have been tested for replacement. The success of some biological and synthetic nanostructures such as cell-penetrating peptides and carbon nanotubes in transferring macromolecules (proteins and nucleic acids) into mammalian cells provoked us to assess the ability of an engineered chimeric peptide and also arginine functionalized single-walled carbon nanotube in gene delivery to intact tobacco (Nicotiana tabacum var...
December 2018: Molecular Biotechnology
Sébastien Sart, Spiros N Agathos
Mesenchymal stem cells and pluripotent stem cells are recognized as promising tools for tissue engineering, cell therapy, and drug screening. Their use in therapy requires the production of a sufficient number of cells committed to functional regenerative phenotypes. Time- and magnitude-controlled application of mechanical and biochemical cues is required to appropriately control the evolution of stem cell phenotype in 3D. The temporal monitoring of the impact of these cues on the diverse fates of individual stem cells is also needed to ensure the reliability of the differentiation processes...
November 2018: Molecular Biotechnology
Saeed Khalili, Mohamad Javad Rasaee, Taravat Bamdad, Maysam Mard-Soltani, Majid Asadi Ghalehni, Abolfazl Jahangiri, Mohammad Hassan Pouriayevali, Mohammad Reza Aghasadeghi, Fatemeh Malaei
Nucleic acid immunization has recently exhibited a great promise for immunotherapy of various diseases. However, it is now clear that powerful strategies are imminently needed to improve their efficiency. In this regard, whole bacteriophage particles have been described as efficient DNA vaccine delivery vehicles, capable of circumventing the limitations of naked DNA immunization. Moreover, phage particles could be engineered to display specific peptides on their surfaces. Given these inherent characteristics of phages, we have designed a novel hybrid phage-DNA immunization vector using both M13 and pAAV plasmid elements...
November 2018: Molecular Biotechnology
Monika M Golas, Sakthidasan Jayaprakash, Le T M Le, Zongpei Zhao, Violeta Heras Huertas, Ida S Jensen, Juan Yuan, Bjoern Sander
The human tumor suppressor SMARCB1/INI1/SNF5/BAF47 (SNF5) is a core subunit of the multi-subunit ATP-dependent chromatin remodeling complex SWI/SNF, also known as Brahma/Brahma-related gene 1 (BRM/BRG1)-associated factor (BAF). Experimental studies of SWI/SNF are currently considerably limited by the low cellular abundance of this complex; thus, recombinant protein production represents a key to obtain the SWI/SNF proteins for molecular and structural studies. While the expression of mammalian proteins in bacteria is often difficult, the baculovirus/insect cell expression system can overcome limitations of prokaryotic expression systems and facilitate the co-expression of multiple proteins...
November 2018: Molecular Biotechnology
Wojciech Potocki, Alessandro Negri, Grażyna Peszyńska-Sularz, Krzysztof Hinc, Michał Obuchowski, Adam Iwanicki
Mucosal immunizations are convenient ways of vaccination, which do not require any trained personnel for administration. One of the major challenges for developing an effective mucosal vaccine is finding appropriate adjuvant. Bacillus subtilis endospores have been shown to help solving these obstacles while serving as a platform for presentation of both, antigens and adjuvants. In this study, we have successfully designed and constructed recombinant spores displaying an antigen/adjuvant chimeric protein. We have used a fragment of Clostridium difficile flagellar cap FliD protein as antigen and VQGEESNDK peptide, a fragment of human IL-1β, as adjuvant...
November 2018: Molecular Biotechnology
Rahul Vasudeo Ramekar, Kyong-Cheul Park, Kyu Jin Sa, Ju Kyong Lee
Transposable elements account for up to 85% of the maize genome and have significant implications in crop-improvement and evolutionary analyses. The Mutator (Mu) transposon superfamily, a class of DNA transposons, comprises the most complex and active elements in the maize genome, suggesting a special role in plant evolution. Here, we designed a set of Mu-specific primers based on terminal invert repeats and used a transposon display (TD) method for genotyping. We analyzed the distribution pattern of Mu insertions in teosinte (wild relative), sorghum (distant relative), and domesticated maize accessions (dent, sweet, and waxy)...
November 2018: Molecular Biotechnology
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