Journal, genetic engineering & biotechnology最新文献

{"title":"Correction to: New insights on key genes involved in drought stress response of barley: gene networks reconstruction, hub, and promoter analysis.","authors":"Seyedeh Mehri Javadi, Zahra-Sadat Shobbar, Asa Ebrahimi, Maryam Shahbazi","doi":"10.1186/s43141-021-00158-7","DOIUrl":"https://doi.org/10.1186/s43141-021-00158-7","url":null,"abstract":"","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"19 1","pages":"78"},"PeriodicalIF":0.0,"publicationDate":"2021-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s43141-021-00158-7","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38959318","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The challenge of prognostic markers in acute pancreatitis: internist's point of view.","authors":"Ombretta Para,&nbsp;Lorenzo Caruso,&nbsp;Maria Teresa Savo,&nbsp;Elisa Antonielli,&nbsp;Eleonora Blasi,&nbsp;Fabio Capello,&nbsp;Tiziana Ciarambino,&nbsp;Lorenzo Corbo,&nbsp;Armando Curto,&nbsp;Margherita Giampieri,&nbsp;Lucia Maddaluni,&nbsp;Giacomo Zaccagnini,&nbsp;Carlo Nozzoli","doi":"10.1186/s43141-021-00178-3","DOIUrl":"https://doi.org/10.1186/s43141-021-00178-3","url":null,"abstract":"<p><p>Acute pancreatitis, the most frequent hospitalization reason in internal medicine ward among gastrointestinal diseases, is burdened by high mortality rate. The disease manifests mainly in a mild form, but about 20-30% patients have a severe progress that requires intensive care. Patients presenting with acute pancreatitis should be clinically evaluated for organ failure signs and symptoms. Stratifying patients in the first days from symptoms onset is essential to determine therapy and care setting. The aim of our study is to evaluate prognostic factors for acute pancreatitis patients, hospitalized in internal medicine wards, and moreover, understanding the role of various prognostic scores validated in intensive care setting in predicting in-hospital mortality and/or admission to intensive care unit. We conducted a retrospective study enrolling all patients with diagnosis of acute pancreatitis admitted took an internal medicine ward between January 2013 and May 2019. Adverse outcome was considered in-hospital mortality and/or admission to intensive care unit. In total, 146 patients (137 with positive outcome and 9 with adverse outcome) were enrolled. The median age was (67.89 ± 16.44), with a slight prevalence of male (55.1%) compared to female (44.9%). C protein reactive (p = 0.02), creatinine (p = 0.01), sodium (p = 0.05), and troponin I (p = 0.013) after 48 h were significantly increased in patients with adverse outcome. In our study, progression in SOFA score independently increases the probability of adverse outcome in patients hospitalized with acute pancreatitis. SOFA score > 5 is highly predictive of in-hospital mortality (O.R. 32.00; C.I. 6.73-152.5; p = 0.001) compared to other scores. The use of an easy tool, validated in intensive care setting such as SOFA score, might help to better stratify the risk of in-hospital mortality and/or clinical worsening in patients hospitalized with acute pancreatitis in internal medicine ward.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"19 1","pages":"77"},"PeriodicalIF":0.0,"publicationDate":"2021-05-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s43141-021-00178-3","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"39018928","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Synthesis and characterizations iron oxide carbon nanotubes nanocomposite by laser ablation for anti-microbial applications.","authors":"Moatasem Al-Salih,&nbsp;Syakirah Samsudin,&nbsp;Siti Suri Arshad","doi":"10.1186/s43141-021-00161-y","DOIUrl":"https://doi.org/10.1186/s43141-021-00161-y","url":null,"abstract":"<p><strong>Background: </strong>Environmental contamination by microbes is a major public health concern. A damp environment is one of the potential sources for microbe proliferation. Smart synthesis nanocatalytic coatings on surfaces, food, and material from different pathogen bacteria can inhibit using the Fe₃O₄/CNTs as anti-microbial growth can effectively curb this growing threat. In this present work, the anti-microbial efficacy of synthesis of a compound nanoparticle-containing iron oxide-multi-walled carbon nanotube was combined by laser ablation PLAL and explored the anti-bacterial action of colloidal solution of Fe₃O₄/CNTs NPs that was evaluated against bacteria which is classified as gram-negative (Escherichia coli (E. coli), Klebsiella pneumonia (K. pneumonia), and also that is identified as gram-positive (Streptococcus pyogenes (S .pyogenes) and Staphylococcus aureus (S. aureus) under visible light irradiation.</p><p><strong>Results: </strong>Doping of a minute fraction of iron(III) salt (0.5 mol%) in a volatile solvent (ethanol) was carried out via the sol-gel technique. Fe₃O₄ was further calcined at various temperatures (in the range of 500-700 °C) to evaluate the thermal stability of the Fe₃O₄ nanoporous oxidizer nanoparticles. The physicochemical properties of the samples were characterized through X-ray diffraction (XRD), atomic force microscopy (AFM), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), and UV-Visible spectroscopy techniques. XRD results revealed that the nanoparticles framework of Fe₃O₄ was maintained well up to 650 °C by the Fe dopant. UV-Vis results suggested that absorption property of combination Fe₃O₄/CNTs nanopowder by PLAL was enhanced and the band gap is reduced into 2.0 eV.</p><p><strong>Conclusions: </strong>Density functional theory (DFT) studies emphasize the introduction of Fe+ and Fe2+ ions by replacing other ions in the CNT lattice, therefore creating oxygen vacancies. These further promoted anti-microbial efficiency. A significantly high bacterial inactivation that indicates results was evaluated and that the mean estimations of restraint were determined from triple assessment in every appraisal at 400 ml which represent the best anti-bacterial action against gram-positive and gram-negative microbes.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"19 1","pages":"76"},"PeriodicalIF":0.0,"publicationDate":"2021-05-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8131414/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38994480","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Green synthesis of silk sericin-embedded silver nanoparticles and their antibacterial application against multidrug-resistant pathogens.","authors":"Md Abdullah Al Masud, Hamid Shaikh, Md Shamsul Alam, M Minnatul Karim, M Abdul Momin, M Ariful Islam, G M Arifuzzaman Khan","doi":"10.1186/s43141-021-00176-5","DOIUrl":"10.1186/s43141-021-00176-5","url":null,"abstract":"<p><strong>Background: </strong>The green synthesis strategy of metallic nanoparticles (NPs) has become popular due to being environmentally friendly. Stable silver nanoparticles (AgNPs) have been synthesized by natural products such as starch, soy protein, various extract of leaves, barks, and roots functioning both as reducing and stabilizing agents. Likewise, silk sericin (SS) is a globular protein discarded in the silk factory might be used for NP synthesis. In this research, we focus on the green synthesis and stabilization of AgNPs by SS as well as assessment of their antibacterial activities against some drug-resistant pathogen.</p><p><strong>Results: </strong>SS was extracted from Bombyx mori silkworm cocoons in an aqueous medium. 17 w/w% of dry sericin powder with respect to the cocoon's weight was obtained by freeze-drying. Furthermore, AgNPs conjugated to sericin, i.e., SS-capped silver nanoparticles (SS-AgNPs) were synthesized by easy, cost-effective, and environment-friendly methods. The synthesized SS-AgNPs were characterized by UV-visible spectroscopy, Fourier-transform infrared-attenuated total reflection (FTIR-ATR) spectroscopy, transmission electron microscopy (TEM), and X-ray diffraction measurement. It has been found from the absorbance of UV-visible spectroscopy that a higher percent of SS-AgNPs was obtained at a higher concentration of silver nitrate solution. FTIR-ATR spectra showed that the carboxylate groups obtained from silk sericin act as a reducing agent for the synthesis of silver nanoparticles, while NH₂+ and COO- act as a stabilizer of AgNPs. The X-ray diffractogram of SS-AgNPs was quite different from AgNO₃ and sericin due to a change in the crystal structure. The diameter of AgNPs was around 20-70 nm observed using TEM. The synthesized SS-AgNPs exhibited strong antibacterial activity against multidrug-resistant pathogens, Escherichia coli and Pseudomonas aeruginosa. Minimal inhibitory/bactericidal concentrations against E. coli and P. aeruginosa were 20μg/mL.</p><p><strong>Conclusions: </strong>This study encourages the use of Bombyx mori for the ecofriendly synthesis of SS-AgNPs to control multidrug-resistant microorganisms.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"19 1","pages":"74"},"PeriodicalIF":0.0,"publicationDate":"2021-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8128943/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38920632","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Defining the molecular pathology and consequent phenotypes in Egyptian HB patients.","authors":"Ghada Y El-Kamah, Rehab M Mosaad, Mohamed B Taher, Khalda S Amr","doi":"10.1186/s43141-021-00165-8","DOIUrl":"10.1186/s43141-021-00165-8","url":null,"abstract":"<p><strong>Background: </strong>Hemophilia B (HB) (also known as Christmas disease) is a rare X-linked recessive disorder characterized by spontaneous or prolonged hemorrhages caused by mutations in Factor 9 (F9) gene leading to deficient or defective coagulation F9. Our study aimed at identifying the causative mutations within a sample of HB Egyptian patients. The present study comprised clinical data of eleven HB patients descending from six unrelated families and a seventh family including a carrier mother with a history of deceased HB sibling. Sequencing of F9 gene was performed.</p><p><strong>Results: </strong>The study revealed four mutations; two missense NM_000133.3:c.676C>G, (P.Arg226Gly) and NM_000133.3:c.1305T>G, (p.Cys435Trp), and two nonsense mutations NM_000133.3:c.880C>T, (p.Arg294*) and NM_000133.3:c.1150C>T, (p.Arg384*), identified mutations spanned exons 6 and 8 of which a total of three mutations are located in hotspot exon 8 of F9 gene.</p><p><strong>Conclusions: </strong>Reviewing the literature, this is the first molecular analysis of F9 gene in HB Egyptian patients. Consistent genotype/phenotypic severity correlation could be concluded, helping proper genetic counseling and prenatal decision taking.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"19 1","pages":"75"},"PeriodicalIF":0.0,"publicationDate":"2021-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8128942/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38990905","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Study of lipase producing gene in wheat - an in silico approach.","authors":"Shradha Rani, Priya Kumari, Raju Poddar, Soham Chattopadhyay","doi":"10.1186/s43141-021-00150-1","DOIUrl":"10.1186/s43141-021-00150-1","url":null,"abstract":"<p><strong>Background: </strong>Lipases (EC 3.1.1.3) catalyze the hydrolysis of oil into free fatty acids and glycerol forming the 3rd largest group of commercialized enzymes. Plant lipases grab attention recently because of their specificity, less production and purified cost, and easy availability. In silico approach is the first step to identify different genes coding for lipase in a most common indigenous plant, wheat, to explore the possibility of this plant as an alternative source for commercial lipase production. As the hierarchy organization of genes reflects an ancient process of gene duplication and divergence, many of the theoretical and analytical tools of the phylogenetic systematics can be utilized for comparative genomic studies. Also, in addition to experimental identification and characterization of genes, for computational genomic analysis, Arabidopsis has become a popular strategy to identify crop genes which are economically important, as Arabidopsis genes had been well identified and characterized for lipase. A number of articles had been reported in which genes of wheat have shown strong homology with Arabidopsis. The complete genome sequences of rice and Arabidopsis constitute a valuable resource for comparative genome analysis as they are representatives of the two major evolutionary lineages within the angiosperms. Here, in this in silico approach, Arabidopsis and Oryza sativa serve as models for dicotyledonous and monocotyledonous species, respectively, and the genomic sequence data available was used to identify the lipase genes in wheat.</p><p><strong>Results: </strong>In this present study, Ensembl Plants database was explored for lipase producing gene present in wheat genome and 21 genes were screened down as they contain specific domain and motif for lipase (GXSXG). According to the evolutionary analysis, it was found that the gene TraesCS5B02G157100, located in 5B chromosome, has 58.35% sequence similarity with the reported lipase gene of Arabidopsis thaliana and gene TraesCS3A02G463500 located in the 3A chromosome has 51.74% sequence similarity with the reported lipase gene of Oryza sativa. Homology modeling was performed using protein sequences coded by aforementioned genes and optimized by molecular dynamic simulations. Further with the help of molecular docking of modeled structures with tributyrin, binding efficiency was checked, and the difference in energies (DE) was -9.83 kcal/mol and -6.67 kcal/mol, respectively.</p><p><strong>Conclusions: </strong>The present work provides a basic understanding of the gene-encoding lipase in wheat, which could be easily accessible and used as a potent industrial enzyme. The study enlightens another direction which can be used further to explore plant lipases.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"19 1","pages":"73"},"PeriodicalIF":0.0,"publicationDate":"2021-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8128969/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38908712","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Molecular characterization of a novel β-defensin isoform from the red-toothed trigger fish, Odonus niger (Ruppel, 1836).","authors":"S Neelima,&nbsp;K Archana,&nbsp;P P Athira,&nbsp;M V Anju,&nbsp;V V Anooja,&nbsp;I S Bright Singh,&nbsp;Rosamma Philip","doi":"10.1186/s43141-021-00175-6","DOIUrl":"https://doi.org/10.1186/s43141-021-00175-6","url":null,"abstract":"<p><strong>Background: </strong>The concern regarding a post-antibiotic era with increasing drug resistance by pathogens imposes the need to discover alternatives for existing antibiotics. Antimicrobial peptides (AMPs) with their versatile therapeutic properties are a group of promising molecules with curative potentials. These evolutionarily conserved molecules play important roles in the innate immune system of several organisms. The β-defensins are a group of cysteine rich cationic antimicrobial peptides that play an important role in the innate immune system by their antimicrobial activity against the invading pathogens. The present study deals with a novel β-defensin isoform from the red-toothed trigger fish, Odonus niger. Total RNA was isolated from the gills, cDNA was synthesized and the β-defensin isoform obtained by polymerase chain reaction was cloned and subjected to structural and functional characterization in silico.</p><p><strong>Results: </strong>A β-defensin isoform could be detected from the gill mRNA of red-toothed trigger fish, Odonus niger. The cDNA encoded a 63 amino acid peptide, β-defensin, with a 20 amino acid signal sequence followed by 43 amino acid cationic mature peptide (On-Def) having a molecular weight of 5.214 kDa and theoretical pI of 8.89. On-Def possessed six highly conserved cysteine residues forming disulfide bonds between C1-C5, C2-C4, and C3-C6, typical of β-defensins. An anionic pro-region was observed prior to the β-defensin domain within the mature peptide. Clustal alignment and phylogenetic analyses revealed On-Def as a group 2 β-defensin. Furthermore, it shared some structural similarities and functional motifs with β-defensins from other organisms. On-Def was predicted to be non-hemolytic with anti-bacterial, anti-viral, anti-fungal, anti-cancer, and immunomodulatory potential.</p><p><strong>Conclusion: </strong>On-Def is the first report of a β-defensin from the red-toothed trigger fish, Odonus niger. The antimicrobial profile showed the potential for further studies as a suitable candidate for antimicrobial peptide therapeutics.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"19 1","pages":"71"},"PeriodicalIF":0.0,"publicationDate":"2021-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s43141-021-00175-6","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38973666","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Secondary metabolites and biodiversity of actinomycetes.","authors":"Manal Selim Mohamed Selim, Sayeda Abdelrazek Abdelhamid, Sahar Saleh Mohamed","doi":"10.1186/s43141-021-00156-9","DOIUrl":"10.1186/s43141-021-00156-9","url":null,"abstract":"<p><strong>Background: </strong>The ability to produce microbial bioactive compounds makes actinobacteria one of the most explored microbes among prokaryotes. The secondary metabolites of actinobacteria are known for their role in various physiological, cellular, and biological processes.</p><p><strong>Main body: </strong>Actinomycetes are widely distributed in natural ecosystem habitats such as soil, rhizosphere soil, actinmycorrhizal plants, hypersaline soil, limestone, freshwater, marine, sponges, volcanic cave-hot spot, desert, air, insects gut, earthworm castings, goat feces, and endophytic actinomycetes. The most important features of microbial bioactive compounds are that they have specific microbial producers: their diverse bioactivities and their unique chemical structures. Actinomycetes represent a source of biologically active secondary metabolites like antibiotics, biopesticide agents, plant growth hormones, antitumor compounds, antiviral agents, pharmacological compounds, pigments, enzymes, enzyme inhibitors, anti-inflammatory compounds, single-cell protein feed, and biosurfactant.</p><p><strong>Short conclusions: </strong>Further highlight that compounds derived from actinobacteria can be applied in a wide range of industrial applications in biomedicines and the ecological habitat is under-explored and yet to be investigated for unknown, rare actinomycetes diversity.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"19 1","pages":"72"},"PeriodicalIF":0.0,"publicationDate":"2021-05-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s43141-021-00156-9","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38987550","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Plant regeneration and transformation of Trachyspermum ammi using Agrobacterium tumefaciens and zygotic embryos.","authors":"Masoumeh Nomani,&nbsp;Masoud Tohidfar","doi":"10.1186/s43141-021-00173-8","DOIUrl":"https://doi.org/10.1186/s43141-021-00173-8","url":null,"abstract":"<p><strong>Background: </strong>Trachyspermum ammi is one of the key medicinal plant species with many beneficial properties. Thymol is the most important substance in the essential oil of this plant. Thymol is a natural monoterpene phenol with high anti-microbial, anti-bacterial, and anti-oxidant properties. Thymol in the latest research has a significant impact on slowing the progression of cancer cells in human. In this research, embryos were employed as convenient explants for the fast and effectual regeneration and transformation of T. ammi. To regenerate this plant, Murashige and Skoog (MS) and Gamborg's B5 (B5) media were supplemented with diverse concentrations of plant growth regulators, such as 6-benzyladenine (BA), 1-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), and kinetin (kin). Transgenic Trachyspermum ammi plants were also obtained using Agrobacterium-mediated transformation and zygotic embryos explants. Moreover, two Agrobacterium tumefaciens strains (EHA101 and LBA4404) harboring pBI121-TPS2 were utilized for genetic transformation to Trachyspermum ammi.</p><p><strong>Results: </strong>According to the obtained results, the highest plant-regeneration frequency was obtained with B5 medium supplemented with 0.5 mg/l BA and 1 mg/l NAA. The integrated gene was also approved using the PCR reaction and the Southern blot method. Results also showed that the EHA101 strain outperformed another strain in inoculation time (30 s) and co-cultivation period (1 day) (transformation efficiency 19.29%). Furthermore, HPLC method demonstrated that the transformed plants contained a higher thymol level than non-transformed plants.</p><p><strong>Conclusions: </strong>In this research, a fast protocol was introduced for the regeneration and transformation of Trachyspermum ammi, using zygotic embryo explants in 25-35 days. Our findings confirmed the increase in the thymol in the aerial part of Trachyspermum ammi. We further presented an efficacious technique for enhancing thymol content in Trachyspermum ammi using Agrobacterium-mediated plant transformation system that can be beneficial in genetic transformation and other plant biotechnology techniques.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"19 1","pages":"68"},"PeriodicalIF":0.0,"publicationDate":"2021-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1186/s43141-021-00173-8","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38970525","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Production of polyclonal antibody against the recombinant PirB^vp protein of Vibrio parahaemolyticus.","authors":"Ngoc-Diem Duong, Khai-Hoan Nguyen-Phuoc, Kim-Yen Thi Do, Nguyet-Thu Thi Nguyen, Thuoc Linh Tran, Hieu Tran-Van","doi":"10.1186/s43141-021-00172-9","DOIUrl":"10.1186/s43141-021-00172-9","url":null,"abstract":"<p><strong>Background: </strong>Acute hepatopancreatic necrosis disease (AHPND) is caused by toxin-producing strains of Vibrio parahaemolyticus which contain deadly binary toxins PirA^vp and PirB^vp encoded in pVA1 plasmid. The polyclonal antibodies against PirB^vp protein could be used to develop immunochromatographic test strip for in-field diagnosis of AHPND.</p><p><strong>Results: </strong>In this study, PirB^vp gene was amplified, cloned, and expressed in E. coli. The expressed protein was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot probed with 6xHis antibodies. Then, the recombinant PirB^vp (rPirB^vp) was purified using Ni-Sepharose column. Rabbits were immunized with the purified rPirB^vp, and produced antibodies were analyzed using Ouchterlony double immunodiffusion. The antibody titration and antibody purification were performed by ELISA and affinity chromatography, respectively. Finally, antibody specificity and sensitivity were evaluated by dot blotting. The present study showed a high titer of polyclonal antibodies in rabbit serum after immunization and the titer increased steadily during the immunization schedule. The highest titer of antibody reached up to 2,560,000 with LOD of 0.1 ng/mL. The purified antibodies showed no cross-reactivity with proteins from other Vibrio species, and the detection threshold ranged from 6.25 to 12.5 ng toxin/dot.</p><p><strong>Conclusion: </strong>This study highlights the production of high titer and specific polyclonal antibodies as an initial material towards the development of lateral-flow strip test.</p>","PeriodicalId":74026,"journal":{"name":"Journal, genetic engineering & biotechnology","volume":"19 1","pages":"70"},"PeriodicalIF":0.0,"publicationDate":"2021-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8113428/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"38972725","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}