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https://www.readbyqxmd.com/read/28316015/expression-and-characterization-of-codon-optimized-crimean-congo-hemorrhagic-fever-virus-gn-glycoprotein-in-insect-cells
#1
Mehdi Rahpeyma, Alireza Samarbaf-Zadeh, Manoochehr Makvandi, Ata A Ghadiri, Stuart D Dowall, Fatemeh Fotouhi
Crimean-Congo hemorrhagic fever virus (CCHFV) is a major cause of tick-borne viral hemorrhagic disease in the world. Despite of its importance as a deadly pathogen, there is currently no licensed vaccine against CCHF disease. The attachment glycoprotein of CCHFV (Gn) is a potentially important target for protective antiviral immune responses. To characterize the expression of recombinant CCHFV Gn in an insect-cell-based system, we developed a gene expression system expressing the full-length coding sequence under a polyhedron promoter in Sf9 cells using recombinant baculovirus...
March 18, 2017: Archives of Virology
https://www.readbyqxmd.com/read/28300184/corrigendum-high-level-expression-of-improved-thermo-stable-alkaline-xylanase-variant-in-pichia-pastoris-through-codon-optimization-multiple-gene-insertion-and-high-density-fermentation
#2
Yihong Lu, Cheng Fang, Qinhong Wang, Yuling Zhou, Guimin Zhang, Yanhe Ma
No abstract text is available yet for this article.
March 16, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28296951/in-silico-design-expression-and-purification-of-novel-chimeric-escherichia-coli-o157-h7-ompa-fused-to-ltb-protein-in-escherichia-coli
#3
Aytak Novinrooz, Taghi Zahraei Salehi, Roya Firouzi, Sina Arabshahi, Abdollah Derakhshandeh
E. coli O157:H7, one of the major EHEC serotypes, is capable of developing bloody diarrhea, hemorrhagic colitis (HC), and fatal hemolytic uremic syndrome (HUS) and is accompanied by high annual economic loss worldwide. Due to the increased risk of HC and HUS development following antibiotic therapy, the prevention of infections caused by this pathogen is considered to be one of the most effective ways of avoiding the consequences of this infection. The main aim of the present study was to design, express, and purify a novel chimeric protein to develope human vaccine candidate against E...
2017: PloS One
https://www.readbyqxmd.com/read/28289571/differential-bicodon-usage-in-lowly-and-highly-abundant-proteins
#4
Luis A Diambra
Degeneracy in the genetic code implies that different codons can encode the same amino acid. Usage preference of synonymous codons has been observed in all domains of life. There is much evidence suggesting that this bias has a major role on protein elongation rate, contributing to differential expression and to co-translational folding. In addition to codon usage bias, other preference variations have been observed such as codon pairs. In this paper, I report that codon pairs have significant different frequency usage for coding either lowly or highly abundant proteins...
2017: PeerJ
https://www.readbyqxmd.com/read/28284983/systemic-aav-mediated-%C3%AE-sarcoglycan-delivery-targeting-cardiac-and-skeletal-muscle-ameliorates-histological-and-functional-deficits-in-lgmd2e-mice
#5
Eric R Pozsgai, Danielle A Griffin, Kristin N Heller, Jerry R Mendell, Louise R Rodino-Klapac
Limb-girdle muscular dystrophy type 2E (LGMD2E), resulting from mutations in β-sarcoglycan (SGCB), is a progressive dystrophy with deteriorating muscle function, respiratory failure, and cardiomyopathy in 50% or more of LGMD2E patients. SGCB knockout mice share many of the phenotypic deficiencies of LGMD2E patients. To investigate systemic SGCB gene transfer to treat skeletal and cardiac muscle deficits, we designed a self-complementary AAVrh74 vector containing a codon-optimized human SGCB transgene driven by a muscle-specific promoter...
March 8, 2017: Molecular Therapy: the Journal of the American Society of Gene Therapy
https://www.readbyqxmd.com/read/28283349/aav-gene-therapy-corrects-otc-deficiency-and-prevents-liver-fibrosis-in-aged-otc-knock-out-heterozygous-mice
#6
Lili Wang, Peter Bell, Hiroki Morizono, Zhenning He, Elena Pumbo, Hongwei Yu, John White, Mark L Batshaw, James M Wilson
Ornithine transcarbamylase (OTC) deficiency is an X-linked disorder of the urea cycle. Hemizygous males and heterozygous females may experience life-threatening elevations of ammonia in blood and brain, leading to irreversible cognitive impairment, coma, and death. Recent evidence of acute liver failure and fibrosis/cirrhosis is also emerging in OTC-deficient patients. Here, we investigated the long-term consequences of abnormal ureagenesis in female mice heterozygous (Het) for a null mutation in the OTC gene...
March 2, 2017: Molecular Genetics and Metabolism
https://www.readbyqxmd.com/read/28272978/novel-recombinant-dna-vaccine-candidates-for-human-respiratory-syncytial-virus-preclinical-evaluation-of-immunogenicity-and-protection-efficiency
#7
Mohamed A Farrag, Haitham M Amer, Peter Öhlschläger, Maaweya E Hamad, Fahad N Almajhdi
The development of safe and potent vaccines for human respiratory syncytial virus (HRSV) is still a challenge for researchers worldwide. DNA-based immunization is currently a promising approach that has been used to generate human vaccines for different age groups. In this study, novel HRSV DNA vaccine candidates were generated and preclinically tested in BALB/c mice. Three different versions of the codon-optimized HRSV fusion (F) gene were individually cloned into the pPOE vector. The new recombinant vectors either express full-length (pPOE-F), secretory (pPOE-TF), or M282-90 linked (pPOE-FM2) forms of the F protein...
March 8, 2017: Human Vaccines & Immunotherapeutics
https://www.readbyqxmd.com/read/28271668/does-mrna-structure-contain-genetic-information-for-regulating-co-translational-protein-folding
#8
REVIEW
Jian-Rong Yang
Currently many facets of genetic information are ill-defined. In particular, how protein folding is genetically regulated has been a long-standing issue for genetics and protein biology. And a generic mechanistic model with supports of genomic data is still lacking. Recent technological advances have enabled much needed genome-wide experiments. While putting the effect of codon optimality on debate, these studies have supplied mounting evidence suggesting a role of mRNA structure in the regulation of protein folding by modulating translational elongation rate...
January 18, 2017: Zool Res
https://www.readbyqxmd.com/read/28267627/stop-codon-mutagenesis-for-homogenous-expression-of-human-papillomavirus-l1-protein-in-escherichia-coli
#9
Daning Wang, Fei Fan, Zhihai Li, Xinlin Liu, Shuo Song, Shuangping Wei, Maozhou He, Yahua Lin, Zhongyi Li, Minxi Wei, Hai Yu, Ying Gu, Shaowei Li, Ningshao Xia
Human papillomavirus (HPV) is widely accepted to be the major causative pathogen of cervical cancer, warts, and other epithelial tumors. Virus infection and subsequent disease development can be prevented by vaccination with HPV vaccines derived from eukaryotic expression systems. Here, we report the soluble expression of the major capsid protein L1 of HPV31, a dominant carcinogenic HPV genotype, in Escherichia coli. HPV31 L1 protein and its elongated form (L1+) were observed in SDS-PAGE and CE-SDS analysis, generated by the native HPV31 L1 gene with a TAA stop codon...
March 3, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28262014/high-efficiency-secretion-of-%C3%AE-mannanase-in-bacillus-subtilis-through-protein-synthesis-and-secretion-optimization
#10
Yafeng Song, Gang Fu, Huina Dong, Jianjun Li, Yuguang Du, Dawei Zhang
The manno endo-1, 4-mannosidase (β-mannanase, EC. 3.2.1.78) catalyzes the random hydrolysis of internal (1→4)-β-mannosidic linkages in the mannan polymers. A codon optimized β-mannanase gene from Bacillus licheniformis DSM13 was expressed in Bacillus subtilis. When four Sec-dependent and two Tat-dependent signal peptide sequences cloned from B. subtilis were placed upstream of the target gene, the highest activity of β-mannanase was observed using SPlipA as a signal peptide. Then, a 1.25-fold activity of β-mannanase was obtained when another copy of groESL operon was inserted into the genome of host strain...
March 6, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28260621/differential-regulation-of-hepatitis-b-virus-core-protein-expression-and-genome-replication-by-a-small-upstream-open-reading-frame-and-naturally-occurring-mutations-in-the-precore-region
#11
Li Zong, Yanli Qin, Haodi Jia, Lei Ye, Yongxiang Wang, Jiming Zhang, Jack R Wands, Shuping Tong, Jisu Li
Hepatitis B virus (HBV) transcribes two subsets of 3.5-kb RNAs: precore RNA for hepatitis B e antigen (HBeAg) expression, and pregenomic RNA for core and P protein translation as well as genome replication. HBeAg expression could be prevented by mutations in the precore region, while an upstream open reading frame (uORF) has been proposed as a negative regulator of core protein translation. We employed replication competent HBV DNA constructs and transient transfection experiments in Huh7 cells to verify the uORF effect and to explore the alternative function of precore RNA...
March 2, 2017: Virology
https://www.readbyqxmd.com/read/28254397/high-level-expression-of-two-thermophilic-%C3%AE-mannanases-in-yarrowialipolytica
#12
Wang YaPing, Rao Ben, Zhang Ling, Ma Lixin
Two thermophilic β-mannanases (ManA and ManB)were successfully expressed in Yarrowialipolytica using vector pINA1296I. The sequences of manA from Aspergillus niger CBS 513.88 and manB from Bacillus subtilis BCC41051 were optimized based on codon-usage bias in Y.lipolytica and synthesized by overlapping polymerase chain reaction (PCR). We utilized the pINA1296I vector, which allows inserting and expression of multiple copies of an expression cassette, to engineer recombinant strains containing multiple copies of manA or manB...
February 22, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28251390/co-expression-of-l-glutamate-oxidase-and-catalase-in-escherichia-coli-to-produce-%C3%AE-ketoglutaric-acid-by-whole-cell-biocatalyst
#13
Qingdai Liu, Xiaoqian Ma, Haijiao Cheng, Ning Xu, Jun Liu, Yanhe Ma
OBJECTIVES: To improve the production of α-ketoglutaric acid (α-KG) from L-glutamate by whole-cell biocatalysis. RESULTS: A novel and highly active L-glutamate oxidase, SmlGOX, from Streptomyces mobaraensis was overexpressed and purified. The recombinant SmlGOX was approx. 64 kDa by SDS-PAGE. SmlGOX had a maximal activity of 125 ± 2.7 U mg(-1) at pH 6.0, 35 (o)C. The apparent Km and Vmax values of SmlGOX were 9.3 ± 0.5 mM and 159 ± 3 U mg(-1), respectively...
March 1, 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/28250127/attenuated-human-parainfluenza-virus-type-1-expressing-ebola-virus-glycoprotein-gp-administered-intranasally-is-immunogenic-in-african-green-monkeys
#14
Matthias Lingemann, Xueqiao Liu, Sonja Surman, Bo Liang, Richard Herbert, Ashley D Hackenberg, Ursula J Buchholz, Peter L Collins, Shirin Munir
The recent 2014-2016 Ebola virus (EBOV) outbreak prompted increased efforts to develop vaccines against EBOV disease. We describe the development and preclinical evaluation of an attenuated recombinant human parainfluenza virus type 1 (rHPIV1) expressing the membrane-anchored form of EBOV glycoprotein GP, as an intranasal (IN) EBOV vaccine. GP was codon optimized and expressed either as a full-length protein or an engineered chimeric form in which its transmembrane and cytoplasmic tail (TMCT) domains were substituted with those of the HPIV1 F protein in an effort to enhance packaging into the vector particle and immunogenicity...
March 1, 2017: Journal of Virology
https://www.readbyqxmd.com/read/28249562/nucamino-a-nucleotide-to-amino-acid-alignment-optimized-for-virus-gene-sequences
#15
Philip L Tzou, Xiaoqiu Huang, Robert W Shafer
BACKGROUND: Current nucleotide-to-amino acid alignment software programs were developed primarily for detecting gene exons within eukaryotic genomes and were therefore optimized for speed across long genetic sequences. We developed a nucleotide-to-amino acid alignment program NucAmino optimized for virus sequencing. RESULTS: NucAmino is an open source program written in the high-level language Go. NucAmino is more likely to align codons flush with a reference sequence's amino acids and can be modified to facilitate the placement of insertions and deletions at specific positions...
March 1, 2017: BMC Bioinformatics
https://www.readbyqxmd.com/read/28247198/development-and-application-of-a-t7-rna-polymerase-dependent-expression-system-for-antibiotic-production-improvement-in-streptomyces
#16
Junhong Wei, Jinjin Tian, Guoqing Pan, Jie Xie, Jialing Bao, Zeyang Zhou
OBJECTIVE: To develop a reliable and easy to use expression system for antibiotic production improvement of Streptomyces. RESULTS: A two-compound T7 RNA polymerase-dependent gene expression system was developed to fulfill this demand. In this system, the T7 RNA polymerase coding sequence was optimized based on the codon usage of Streptomyces coelicolor. To evaluate the functionality of this system, we constructed an activator gene overexpression strain for enhancement of actinorhodin production...
February 28, 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/28242529/identification-of-the-key-amino-acid-sites-of-the-carbendazim-hydrolase-mhei-from-a-novel-carbendazim-degrading-strain-mycobacterium-sp-sd-4
#17
Yingkun Zhang, Hui Wang, Xiang Wang, Bo Hu, Chenfei Zhang, Wen Jin, Shijun Zhu, Gang Hu, Qing Hong
A novel carbendazim (methyl-1H-benzimidazol-2-ylcarbamate, or MBC) degrading strain SD-4 was isolated and identified preliminarily as Mycobacterium sp. according to its phenotypic features and phylogenetic analysis. This strain could utilize MBC as the sole carbon and nitrogen sources for growth and degrade 50mgL(-1) MBC at the average degradation rate of 0.63mgL(-1)h(-1). Strain SD-4 degraded MBC through the typical pathway, in which MBC was first hydrolyzed by MheI to 2-aminobenzimidazole (2-AB) and then converted to 2-hydroxybenzimidazole (2-HB)...
February 16, 2017: Journal of Hazardous Materials
https://www.readbyqxmd.com/read/28238783/visualization-of-glucagon-secretion-from-pancreatic-%C3%AE-cells-by-bioluminescence-video-microscopy-identification-of-secretion-sites-in-the-intercellular-contact-regions
#18
Satoru Yokawa, Takahiro Suzuki, Satoshi Inouye, Yoshikazu Inoh, Ryo Suzuki, Takao Kanamori, Tadahide Furuno, Naohide Hirashima
We have firstly visualized glucagon secretion using a method of video-rate bioluminescence imaging. The fusion protein of proglucagon and Gaussia luciferase (PGCG-GLase) was used as a reporter to detect glucagon secretion and was efficiently expressed in mouse pancreatic α cells (αTC1.6) using a preferred human codon-optimized gene. In the culture medium of the cells expressing PGCG-GLase, luminescence activity determined with a luminometer was increased with low glucose stimulation and KCl-induced depolarization, as observed for glucagon secretion...
April 15, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28238082/a-novel-bicistronic-expression-system-composed-of-the-intraflagellar-transport-protein-gene-ift25-and-fmdv-2a-sequence-directs-robust-nuclear-gene-expression-in-chlamydomonas-reinhardtii
#19
Bin Dong, He-He Hu, Zhen-Fang Li, Rong-Qiang Cheng, De-Mei Meng, Junping Wang, Zhen-Chuan Fan
Chlamydomonas reinhardtii offers a great promise for large-scale production of multiple recombinant proteins of pharmaceutical and industrial interest. However, the nuclear-encoding transgenes usually are expressed at a low level, which severely hampers the use of this alga in molecular farming. In this study, the promoter of the endogenous intraflagellar transport 25 (IFT25) gene of C. reinhardtii was tested for its ability to drive the expression of green fluorescent protein (GFP), which functions as a readout for target gene expression...
February 25, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28229907/expression-and-functional-evaluation-of-biopharmaceuticals-made-in-plant-chloroplasts
#20
REVIEW
Bei Zhang, Balamurugan Shanmugaraj, Henry Daniell
After approval of the first plant-made biopharmaceutical by FDA for human use, many protein drugs are now in clinical development. Within the last decade, significant advances have been made in expression of heterologous complex/large proteins in chloroplasts of edible plants using codon optimized human or viral genes. Furthermore, advances in quantification enable determination of in-planta drug dosage. Oral delivery of plastid-made biopharmaceuticals (PMB) is affordable because it eliminates prohibitively expensive fermentation, purification processes addressing major challenges of short shelf-life after cold storage...
February 21, 2017: Current Opinion in Chemical Biology
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