nucleotide sequence

  • 文章类型: Journal Article
    背景:随着生物数据和计算能力的指数增长,熟悉生物信息学已成为学术界和工业界的一项苛刻和流行的技能。有必要提高学生的能力,以便能够从事生物信息学职业,为了让他们熟悉数据科学领域的科学专业以及追求这些专业所需的学术培训,在一个需求大于供应的领域。
    方法:在这里,我们将一组生物信息学活动实施到研究生课程的蛋白质结构和功能课程中。简洁地说,通过一个为期一学期的案例研究,作为基于探究的生物信息学练习,为学生提供了实践机会,探索基于生物信息学的生物分子数据和结构生物学分析。在学期结束时,学生们还设计并提出了一个作业项目,使他们能够记录他们在学期中使用生物信息学知识鉴定的未知蛋白质。
    结果:模块后的调查反应和学生在实验室模块中的表现意味着它进一步加深了对生物信息学的深入了解。尽管在学习该模块之前没有太多生物信息学的先验知识,但学生表示积极的反馈。
    结论:学生们熟悉了交叉索引数据库,这些数据库将有关蛋白质的重要数据相互联系,酶和基因。这种以研究为基础的生物信息学教学方法磨练的基本技能将使学生能够利用这些知识为他们在生物信息学领域的未来努力。
    BACKGROUND: With an exponential growth in biological data and computing power, familiarity with bioinformatics has become a demanding and popular skill set both in academia and industry. There is a need to increase students\' competencies to be able to take on bioinformatic careers, to get them familiarized with scientific professions in data science and the academic training required to pursue them, in a field where demand outweighs the supply.
    METHODS: Here we implemented a set of bioinformatic activities into a protein structure and function course of a graduate program. Concisely, students were given hands-on opportunities to explore the bioinformatics-based analyses of biomolecular data and structural biology via a semester-long case study structured as inquiry-based bioinformatics exercises. Towards the end of the term, the students also designed and presented an assignment project that allowed them to document the unknown protein that they identified using bioinformatic knowledge during the term.
    RESULTS: The post-module survey responses and students\' performances in the lab module imply that it furthered an in-depth knowledge of bioinformatics. Despite having not much prior knowledge of bioinformatics prior to taking this module students indicated positive feedback.
    CONCLUSIONS: The students got familiar with cross-indexed databases that interlink important data about proteins, enzymes as well as genes. The essential skillsets honed by this research-based bioinformatic pedagogical approach will empower students to be able to leverage this knowledge for their future endeavours in the bioinformatics field.
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  • 文章类型: Journal Article
    在过去的十年里,petabase规模的高通量测序数据的产生和积累带来了巨大的挑战,包括访问人类数据,以及转移,storage,共享大量数据。为了促进数据驱动的生物学研究,韩国政府宣布,政府资助的研究项目产生的所有生物数据应存放在韩国生物数据站(K-BDS),它由多个用于各个数据类型的数据库组成。这里,我们介绍了韩国核苷酸档案(KoNA),核苷酸序列数据的储存库。截至2022年7月,KoNA的韩国阅读档案已从国家基因组项目中收集了超过477TB的原始下一代测序数据。为了确保数据质量并为国际接轨做准备,采用了标准操作程序,这类似于国际核苷酸序列数据库合作。标准操作程序包括使用自动化管道对提交的数据和元数据进行质量控制。其次是手工检查。为了确保快速稳定的数据传输,KoNA中使用了一种称为GBox的高速传输系统。此外,通过GBox上传到KoNA或从KoNA下载的数据可以使用名为Bio-Express的云计算服务轻松处理。这种KoNA的无缝耦合,GBox,Bio-Express增强了数据体验,包括提交,access,和原始核苷酸序列的分析。KoNA不仅满足了韩国国家序列库的未满足需求,而且还为全球研究人员提供数据集,并为基因组学的进步做出了贡献。KoNA可在https://www上获得。Kobic.re.kr/kona/。
    During the last decade, the generation and accumulation of petabase-scale high-throughput sequencing data have resulted in great challenges, including access to human data, as well as transfer, storage, and sharing of enormous amounts of data. To promote data-driven biological research, the Korean government announced that all biological data generated from government-funded research projects should be deposited at the Korea BioData Station (K-BDS), which consists of multiple databases for individual data types. Here, we introduce the Korean Nucleotide Archive (KoNA), a repository of nucleotide sequence data. As of July 2022, the Korean Read Archive in KoNA has collected over 477 TB of raw next-generation sequencing data from national genome projects. To ensure data quality and prepare for international alignment, a standard operating procedure was adopted, which is similar to that of the International Nucleotide Sequence Database Collaboration. The standard operating procedure includes quality control processes for submitted data and metadata using an automated pipeline, followed by manual examination. To ensure fast and stable data transfer, a high-speed transmission system called GBox is used in KoNA. Furthermore, the data uploaded to or downloaded from KoNA through GBox can be readily processed using a cloud computing service called Bio-Express. This seamless coupling of KoNA, GBox, and Bio-Express enhances the data experience, including submission, access, and analysis of raw nucleotide sequences. KoNA not only satisfies the unmet needs for a national sequence repository in Korea but also provides datasets to researchers globally and contributes to advances in genomics. The KoNA is available at https://www.kobic.re.kr/kona/.
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  • 文章类型: Journal Article
    The objective is to determine the complete nucleotide sequence and conduct a phylogenetic analysis of genome variants of the Puumala virus isolated in the Saratov region.
    METHODS: The samples for the study were field material collected in the Gagarinsky (formerly Saratovsky), Engelssky, Novoburassky and Khvalynsky districts of the Saratov region in the period from 2019 to 2022. To specifically enrich the Puumala virus genome in the samples, were used PCR and developed a specific primer panel. Next, the resulting PCR products were sequenced and the fragments were assembled into one sequence for each segment of the virus genome. To construct phylogenetic trees, the maximum parsimony algorithm was used.
    RESULTS: Genetic variants of the Puumala virus isolated in the Saratov region have a high degree of genome similarity to each other, which indicates their unity of origin. According to phylogenetic analysis, they all form a separate branch in the cluster formed by hantaviruses from other subjects of the Volga Federal District. The virus variants from the Republics of Udmurtia and Tatarstan, as well as from the Samara and Ulyanovsk regions, are closest to the samples from the Saratov region.
    CONCLUSIONS: The data obtained show the presence of a pronounced territorial confinement of strains to certain regions or areas that are the natural biotopes of their carriers. This makes it possible to fairly accurately determine the territory of possible infection of patients and/or the circulation of carriers of these virus variants based on the sequence of individual segments of their genome.
    Цель работы – определение полной нуклеотидной последовательности и проведение филогенетического анализа вариантов геномов вируса Пуумала, выделенных на территории Саратовской области. Материалы и методы. Образцами для исследования послужил полевой материал, собранный в Гагаринском (бывшем Саратовском), Энгельсском, Новобурасском и Хвалынском районах Саратовской области в период с 2019 по 2022 г. Для специфического обогащения генома вируса Пуумала в образцах использовали ПЦР и панель праймеров, подготовленную для данного исследования. Далее проводили секвенирование полученных продуктов реакции и сборку фрагментов в одну последовательность для каждого из сегментов генома вируса. При построении филогенетических деревьев применяли алгоритм maximum parsimony. Результаты. Показано, что генетические варианты вируса Пуумала, выделенные в Саратовской области, имеют высокую степень подобия генома, что говорит о единстве их происхождения. По данным филогенетического анализа, все выделенные варианты вируса (за исключением изолятов вируса из Хвалынского района) образуют обособленную ветвь в кластере, сформированном хантавирусами из других субъектов Приволжского федерального округа. Самыми близкими к образцам из Саратовской области являются варианты вируса из республик Удмуртия и Татарстан, а также из Самарской и Ульяновской областей. Заключение. Полученные данные указывают на наличие выраженной территориальной приуроченности штаммов к определенным регионам или областям, являющимся природными биотопами их носителей. Этот факт позволяет довольно точно определять территорию возможного инфицирования заболевших и/или циркуляцию переносчиков данных вариантов вируса по последовательности отдельных сегментов их генома..
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  • 文章类型: English Abstract
    Introduction. During the development of the SARS-CoV-2 pandemic in Antioquia, we experienced epidemiological peaks related to the α, ɣ, β, ƛ, and δ variants. δ had the highest incidence and prevalence. This lineage is of concern due to its clinical manifestations and epidemiological characteristics. A total of 253 δ sublineages have been reported in the PANGOLIN database. The sublineage identification through genomic analysis has made it possible to trace their evolution and propagation. Objective. To characterize the genetic diversity of the different SARS-CoV-2 δ sublineages in Antioquia and to describe its prevalence. Materials and methods. We collected sociodemographic information from 2,675 samples, and obtained 1,115 genomes from the GISAID database between July 12th, 2021, and January 18th, 2022. From the analyzed genomes, 515 were selected because of their high coverage values (>90%) to perform phylogenetic analysis and to infer allele frequencies of mutations of interest. Results. We characterized 24 sublineages. The most prevalent was AY.25. Mutations of interest as L452R, P681R, and P681H were identified in this sublineage, comprising a frequency close to 0.99. Conclusions. This study identified that the AY.25 sublineage has a transmission advantage compared to the other δ sublineages. This attribute may be related to the presence of the L452R and P681R mutations associated in other studies with higher evasion of the immune system and less efficacy of drugs against SARS-CoV-2.
    Introducción. Durante el desarrollo de la pandemia por SARS-CoV-2 en Antioquia se presentaron picos epidemiológicos relacionados con las variantes α, ɣ, β, ƛ y δ, donde δ tuvo la mayor incidencia y prevalencia. Este linaje se considera una variante de preocupación dadas las manifestaciones clínicas que desencadena y sus características epidemiológicas. Se han informado 253 sublinajes δ en la base de datos PANGOLIN. La identificación de estos sublinajes mediante análisis genómico ha permitido rastrear su evolución y propagación. Objetivo. Caracterizar la diversidad genética de los diferentes sublinajes δ de SARSCoV-2 en Antioquia y determinar su prevalencia. Materiales y métodos. Se recopiló información sociodemográfica de 2.675 muestras y de 1.115 genomas del repositorio GISAID entre el 12 de julio de 2021 y el 18 de enero de 2022. Se seleccionaron 501 por su alto porcentaje de cobertura (>90 %) para realizar análisis filogenéticos e inferencia de frecuencias alélicas de mutaciones de interés. Resultados. Se caracterizaron 24 sublinajes donde el más prevalente fue AY.25. En este sublinaje se identificaron mutaciones de interés como L452R, P681R y P681H, que comprendían una frecuencia cercana a 0,99. Conclusiones. Este estudio permitió identificar que el sublinaje AY.25 tiene una ventaja de transmisión en comparación con los otros sublinajes δ. Esto puede estar relacionado con la presencia de las mutaciones L452R y P681R que en otros estudios se han visto asociadas con una mayor transmisibilidad, evasión del sistema inmunitario y menor eficacia de los medicamentos contra SARS-CoV-2.
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  • 文章类型: Journal Article
    背景:1-甲基腺苷(m1A)是甲基腺苷的变体,在第1位具有甲基取代基,在RNA稳定性和人代谢物中具有突出作用。
    目标:传统方法,如质谱和定点诱变,被证明是耗时和复杂的。
    方法:本研究集中于使用新的特征开发机制鉴定RNA序列中的m1A位点。获得的特征用于训练集合模型,包括混合,升压,和装袋。然后对训练的集合模型进行独立测试和k-fold交叉验证。
    结果:所提出的模型优于现有的预测因子,并显示了基于主要准确性指标的优化分数。
    结论:出于研究目的,可以通过https://taseersuleman-m1a-ensem1访问所建议模型的用户友好的网络服务器。流光。app/.
    BACKGROUND: 1-methyladenosine (m1A) is a variant of methyladenosine that holds a methyl substituent in the 1st position having a prominent role in RNA stability and human metabolites.
    OBJECTIVE: Traditional approaches, such as mass spectrometry and site-directed mutagenesis, proved to be time-consuming and complicated.
    METHODS: The present research focused on the identification of m1A sites within RNA sequences using novel feature development mechanisms. The obtained features were used to train the ensemble models, including blending, boosting, and bagging. Independent testing and k-fold cross validation were then performed on the trained ensemble models.
    RESULTS: The proposed model outperformed the preexisting predictors and revealed optimized scores based on major accuracy metrics.
    CONCLUSIONS: For research purpose, a user-friendly webserver of the proposed model can be accessed through https://taseersuleman-m1a-ensem1.streamlit.app/ .
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  • 文章类型: Journal Article
    本文研究的目的是基于确定一组特定基因(基因座)的核苷酸序列,将新引入的乳杆菌菌株系统化。解决此问题的主要方法包括进行实验室实验。在这个实验中,对一组由乳杆菌属的小DNA元素组成的有机化合物进行了彻底的检查。多位点基因分型方法是中心技术,辅以额外的分子生物学和种群方法。这些其他方法用于确定乳杆菌纯培养物之间的系统发育相似性程度,并对其进行相应分类。本文介绍了用于多位点分型的基因分离株;揭示了适合多位点基因分型的干酪乳杆菌分离株的数量;揭示了将干酪乳杆菌分离株分为五种测序类型的基因等位基因;证实了遗传分型方法用于多位点测序的有效性。本文对分子生物学领域的微生物学家和遗传学家具有实用价值,以及食品行业的技术人员。随着遗传系统学应用方法的发展,研究乳杆菌属物种的纯培养已经成为可能。应用现代方法对乳酸菌物种进行基因型分类将有可能提高在食品工业和医学中使用更好,更安全的产品的效率。
    The aim of the study in this article is to systematise the newly introduced strains of Lactobacillus based on determining the nucleotide sequence of a particular set of their genes (loci). The primary approach employed to address this issue involves conducting a laboratory experiment. During this experiment, a thorough examination was carried out on a set of organic compounds consisting of small DNA elements from the Lactobacillus genus. The Multilocus genotyping method served as the central technique, complemented by additional molecular-biological and population methods. These additional methods were utilized to determine the extent of phylogenetic similarity among pure cultures of Lactobacillus and to classify them accordingly. The article presents the gene isolates that were used for Multilocus typing; the number of L. casei isolates suitable for Multilocus genotyping was revealed; the gene alleles that allowed classifying L. casei isolates into five sequencing types were revealed; the effectiveness of genetic typing method for Multilocus sequencing was substantiated. The article is of practical value for microbiologists and geneticists in the field of molecular biology, as well as for technologists in the food industry. With the development of applied methods in genetic systematics, it has become possible to study pure culture of Lactobacillus species. The application of modern methods of genotypic classification of Lactobacillus species will make it possible to increase the efficiency of using better and safer products in the food industry and medicine.
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  • 文章类型: Journal Article
    描述错误鉴定的核苷酸序列试剂的人类基因研究大多在低至中等影响因子的期刊中被鉴定出来,不可靠的发现可能被认为对未来研究的影响有限。这项研究调查了描述错误鉴定的核苷酸序列的论文是否也发表在高影响因子癌症研究期刊上。我们在2014年、2016年、2018年和2020年发表的原始分子癌症文章中手动验证了核苷酸序列同一性,包括声称靶向circRNAs的核苷酸序列试剂。使用一些2018年和2020年分子癌症论文中确定的关键词,我们还在研究miRNA和/或circRNA的2020篇Oncogene论文中验证了核苷酸序列同一性.总的来说,3.8%(251/6647)和4.0%(47/1165)的核苷酸序列在分子癌症和癌基因论文中得到验证,分别,被发现被错误地识别。错误鉴定的核苷酸序列分布在18%(91/500)原始分子癌症论文中,包括38%(31/82)2020年的分子癌症论文,以及40%(21/52)2020年的癌基因论文。因此,具有错误鉴定的核苷酸序列的原始论文在两个高影响因素的癌症研究期刊中出乎意料地频繁出现。强调使用期刊影响因子或引文作为研究质量代理的风险。
    Human gene research studies that describe wrongly identified nucleotide sequence reagents have been mostly identified in journals of low to moderate impact factor, where unreliable findings could be considered to have limited influence on future research. This study examined whether papers describing wrongly identified nucleotide sequences are also published in high-impact-factor cancer research journals. We manually verified nucleotide sequence identities in original Molecular Cancer articles published in 2014, 2016, 2018, and 2020, including nucleotide sequence reagents that were claimed to target circRNAs. Using keywords identified in some 2018 and 2020 Molecular Cancer papers, we also verified nucleotide sequence identities in 2020 Oncogene papers that studied miRNA(s) and/or circRNA(s). Overall, 3.8% (251/6647) and 4.0% (47/1165) nucleotide sequences that were verified in Molecular Cancer and Oncogene papers, respectively, were found to be wrongly identified. Wrongly identified nucleotide sequences were distributed across 18% (91/500) original Molecular Cancer papers, including 38% (31/82) Molecular Cancer papers from 2020, and 40% (21/52) selected Oncogene papers from 2020. Original papers with wrongly identified nucleotide sequences were therefore unexpectedly frequent in two high-impact-factor cancer research journals, highlighting the risks of employing journal impact factors or citations as proxies for research quality.
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  • 文章类型: Journal Article
    背景:弧菌是在地表水中发现的自生细菌种群,与各种危及生命的肠外疾病有关,特别是在患有潜在疾病和伤口感染的人群中。目前,关于这些物种的毒力和抗性基因的突变多样性,存在非常小的信息。这项研究评估了核酸内切酶的变异以及弧菌分离株的毒力和抗性基因的突变多样性,携带毒力相关基因(vcgCPI),二氢蝶呤合酶1型和II型基因(Sul1和11),(aadA)氨基糖苷(3\'\')(9)腺苷酸转移酶基因,(aac(3)-IIa,(aacC2)a,氨基糖苷N(3)-乙酰转移酶III,和(strA)氨基糖苷3'-磷酸转移酶抗性基因。
    方法:使用分子生物学技术的组合,生物信息学工具,和序列分析。
    结果:我们的结果揭示了在核苷酸位置(密码子)73-75(A→G)和300-302(N→S)处,创伤弧菌(vcgCPI)的毒力决定子的各种核苷酸变异。弧菌属的氨基糖苷类抗性基因(aadA)描述了482位的核苷酸差异(A→G),而氨基糖苷类耐药基因(sul1和11)显示两个核苷酸多态性可变区(102和140)。氨基酸差异存在于位置140的核苷酸多态性(A→E)。限制酶HinP1I产生的条带模式,MwoI,和StyD4I显示出显著的变化。此外,蛋白质二氢蝶呤合酶1型和II型基因(Sul1和11)的限制酶消化显着不同,而酶DpnI和Hinf1表示没有显著差异。与来自GenBank的参考分离株相比,限制酶NlaIV没有显示条带。然而,抗性决定簇显示显著的点核苷酸突变,不会产生任何具有不同多态性区域的氨基酸变化,正如限制摘要中所揭示的那样。
    结论:描述的毒力和抗性决定子具有与致病基因组学研究相关的特定多态性位点,药物基因组学,和控制这些与水相关的菌株。
    BACKGROUND: Vibrio species are among the autochthonous bacterial  populations found in surface waters and associated with various life-threatening extraintestinal diseases, especially in human populations with underlying illnesses and wound infections. Presently, very diminutive information exists regarding these species\' mutational diversity of virulence and resistance genes. This study evaluated variations in endonucleases and mutational diversity of the virulence and resistance genes of Vibrio isolates, harboring virulence-correlated gene (vcgCPI), dihydropteroate synthase type 1 and type II genes (Sul 1 and 11), (aadA) aminoglycoside (3\'\') (9) adenylyltransferase gene, (aac(3)-IIa, (aacC2)a, aminoglycoside N(3)-acetyltransferase III, and (strA) aminoglycoside 3\'-phosphotransferase resistance genes.
    METHODS: Using combinations of molecular biology techniques, bioinformatics tools, and sequence analysis.
    RESULTS: Our result revealed various nucleotide variations in virulence determinants of V. vulnificus (vcgCPI) at nucleotide positions (codon) 73-75 (A → G) and 300-302 (N → S). The aminoglycosides resistance gene (aadA) of Vibrio species depicts a nucleotide difference at position 482 (A → G), while the aminoglycosides resistance gene (sul 1 and 11) showed two variable regions of nucleotide polymorphism (102 and 140). The amino acid differences exist with the nucleotide polymorphism at position 140 (A → E). The banding patterns produced by the restriction enzymes HinP1I, MwoI, and StyD4I showed significant variations. Also, the restriction enzyme digestion of protein dihydropteroate synthase type 1 and type II genes (Sul 1 and 11) differed significantly, while enzymes DpnI and Hinf1 indicate no significant differences. The restriction enzyme NlaIV showed no band compared to reference isolates from the GenBank. However, the resistant determinants show significant point nucleotide mutation, which does not produce any amino acid change with diverse polymorphic regions, as revealed in the restriction digest profile.
    CONCLUSIONS: The described virulence and resistance determinants possess specific polymorphic locus relevant to pathogenomics studies, pharmacogenomic, and control of such water-associated strains.
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  • 文章类型: Journal Article
    患有呼吸综合征的幼鸟和成年鸟的传染病由于生产力下降和死亡率显著而对工业家禽养殖的发展具有显著的阻碍作用。对抗病毒性疾病的唯一有效方法是及时和有针对性地接种疫苗,这在很大程度上取决于实验室诊断结果。
    本文旨在研究实时逆转录聚合酶链反应,RT-PCR(RT-PCR)有望更有效地诊断鸡传染性支气管炎和新城疫的疫苗株。
    在相关性病毒感染中鉴别诊断病原体的最快,最准确的方法是RT-PCR。文章中提出的选择引物进行扩增的方法使得有可能使用该方法同时进行两种或多种病毒试剂的种间差异诊断。大大加快了诊断速度。
    从测序获得的核苷酸序列与特定病毒株的相关性由于缺乏用于分离遗传组的单一命名机制而变得复杂。
    这项研究的结果将允许将序列轻松,快速地分型为已知和数据库化的病毒株,并避免将来在遗传群体命名法中进一步混淆。
    Infectious diseases of young and adult birds with respiratory syndrome are a significant deterrent to the development of industrial poultry farming due to decreased productivity and significant mortality. The only effective method of combating viral diseases is timely and targeted vaccination, which largely depends on laboratory diagnostic results.
    This article aims to study the real-time reverse transcription polymerase chain reaction, (RT-PCR) which has the prospect of more effective diagnosis of vaccine strains of chicken infectious bronchitis and Newcastle disease.
    The fastest and most accurate method for the differential diagnosis of pathogens in an associative viral infection is RT-PCR. The method proposed in the article for selecting primers for amplification made it possible to use this method for the simultaneous interspecies differential diagnosis of two or more viral agents, significantly accelerating their diagnosis.
    The correlation of the nucleotide sequence obtained from sequencing to a specific virus strain is complicated by the lack of a single nomenclature mechanism for separating genetic groups.
    The results of this study will allow easy and fast typing of sequences into known and databased virus strains and avoid further confusion in the nomenclature of genetic groups in the future.
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  • 文章类型: Journal Article
    获得了来自肺炎链球菌的肺炎溶血素的重组形式。利用VectorNTIAdvance11.0生物信息学分析软件,设计特异性引物以扩增菌株No.3358肺炎链球菌血清型19F含有编码全长肺炎链球菌溶血素蛋白的核苷酸序列。获得分子量对应于编码全长肺炎链球菌溶血素的肺炎链球菌基因组片段的核苷酸序列的PCR产物。构建了重组肺炎球菌溶血素在大肠杆菌中的表达系统。测序证实了编码在大肠杆菌M15菌株中合成的全长重组肺炎球菌溶血素的插入核苷酸序列的同一性。重组蛋白的纯化通过使用Ni-Sepharose在8M尿素缓冲溶液中的亲和层析进行。通过用针对肺炎球菌溶血素的单克隆抗体进行免疫印迹来确认重组蛋白。
    A recombinant form of pneumolysin from Streptococcus pneumoniae was obtained. By using Vector NTI Advance 11.0 bioinformatic analysis software, specific primers were designed in order to amplify the genome fragment of strain No. 3358 S. pneumoniae serotype 19F containing the nucleotide sequence encoding the full-length pneumolysin protein. A PCR product with a molecular weight corresponding to the nucleotide sequence of the S. pneumoniae genome fragment encoding the full-length pneumolysin was obtained. An expression system for recombinant pneumolysin in E. coli was constructed. Sequencing confirmed the identity of the inserted nucleotide sequence encoding the full-length recombinant pneumolysin synthesized in E. coli M15 strain. Purification of the recombinant protein was performed by affinity chromatography using Ni-Sepharose in 8 M urea buffer solution. Confirmation of the recombinant protein was performed by immunoblotting with monoclonal antibodies to pneumolysin.
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