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Draft Genome Sequences for the Frankia sp. strains CgS1, CcI156 and CgMI4, Nitrogen-Fixing Bacteria Isolated from Casuarina sp. in Egypt. 埃及木麻黄属固氮细菌Frankia sp.菌株CgS1、CcI156和CgMI4的基因组序列草图
Pub Date : 2020-09-23 eCollection Date: 2020-01-01 DOI: 10.7150/jgen.51181
Samira Mansour, Erik Swanson, Céline Pesce, Stephen Simpson, Krystalynne Morris, W Kelley Thomas, Louis S Tisa

Frankia sp. strains CgS1, CcI156 and CgMI4 were isolated from Casuarina glauca and C. cunninghamiana nodules. Here, we report the 5.26-, 5.33- and 5.20-Mbp draft genome sequences of Frankia sp. strains CgS1, CcI156 and CgMI4, respectively. Analysis of the genome revealed the presence of high numbers of secondary metabolic biosynthetic gene clusters.

Frankia sp.菌株CgS1、CcI156和CgMI4分别从木麻黄和cunninghamiana根瘤中分离得到。本文报道了Frankia sp.菌株CgS1、CcI156和CgMI4分别为5.26、5.33和5.20 mbp的基因组草图序列。基因组分析显示存在大量次生代谢生物合成基因簇。
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引用次数: 2
Nucleotide sequencing of the HoxA gene cluster using Gorilla fosmid clones. 大猩猩fosmid克隆HoxA基因簇的核苷酸测序。
Pub Date : 2020-08-29 eCollection Date: 2020-01-01 DOI: 10.7150/jgen.50468
Takashi Kitano, Choong-Gon Kim, Naruya Saitou

We sequenced the western gorilla (Gorilla gorilla) HoxA cluster region using seven fosmid clones, and found that the total tiling path sequence was 214,185 bp from the 5' non-genic region of HoxA1 to the 3' non-genic region of Evx1. We compared the nucleotide sequence with the gorilla genome sequence in the NCBI database, and the overall proportion of nucleotide difference was estimated to be 0.0005-0.0007. These estimates are lower than overall genomic polymorphism in gorillas.

利用7个fosmid克隆对西部大猩猩(gorilla gorilla) HoxA聚类区进行测序,发现从HoxA1的5′非基因区到Evx1的3′非基因区,总平纹路径序列为214,185 bp。我们将该核苷酸序列与NCBI数据库中的大猩猩基因组序列进行比较,估计总核苷酸差异比例为0.0005-0.0007。这些估计值低于大猩猩的整体基因组多态性。
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引用次数: 0
Draft genome sequence of multi-resistant Salmonella enterica subsp. enterica serovar Rissen strain 19CS0416 isolated from Vietnam reveals mcr-1 plasmid mediated resistance to colistin already in 2013. 多重耐药肠沙门氏菌亚种基因组序列草图。2013年越南分离的大肠杆菌血清型Rissen菌株19CS0416已显示出mcr-1质粒介导的对粘菌素的耐药性。
Pub Date : 2020-07-03 eCollection Date: 2020-01-01 DOI: 10.7150/jgen.42790
Belen Gonzalez-Santamarina, Anne Busch, Silvia Garcia-Soto, Mostafa Y Abdel-Glil, Jörg Linde, Reinhard Fries, Diana Meemken, Helmut Hotzel, Herbert Tomaso

We report the first draft genome sequence of a Salmonella strain with plasmid-mediated resistance to colistin encoded by mcr-1 gene in Vietnam. Salmonella enterica subsp. enterica serovar Rissen was isolated from a Vietnamese pig slaughterhouse in 2013. We can confirm that mcr-1 gene is identical to the first reported mcr-1 gene of the Escherichia coli strain SHP45, isolated in 2015 from a Chinese pig. The plasmid containing this gene in the strain 19CS0416 was highly related (96.86% identity) to the plasmid (pHNSHP45) contained in this Chinese strain. Moreover, this plasmid was determined to be 100% identical to a plasmid (p13P477T-7) belonging to an Escherichia coli (13P477T) found in Hong Kong during the same year in pigs. Our results will aid in understanding the dissemination of mcr-1 gene in East Asia, dating back to as early as 2013.

我们报道了越南一株由mcr-1基因编码的质粒介导的粘菌素耐药沙门氏菌的第一个基因组序列草图。肠道沙门氏菌亚种2013年在越南一家生猪屠宰场分离出血清瑞森肠杆菌。我们可以确认mcr-1基因与2015年从中国猪中分离出的大肠杆菌SHP45株的首次报道的mcr-1基因相同。菌株19CS0416中含有该基因的质粒与该菌株的质粒pHNSHP45高度相关(同源性为96.86%)。此外,该质粒与同年在香港猪身上发现的大肠杆菌(13P477T)的质粒(p13P477T-7) 100%相同。我们的研究结果将有助于了解mcr-1基因在东亚的传播,最早可追溯到2013年。
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引用次数: 3
Complete genome and plasmid sequences of a multidrug-resistant Salmonella enterica subsp. enterica serovar Panama isolate from a German cattle farm. 多药耐药肠炎沙门氏菌亚群全基因组和质粒序列。从德国养牛场分离出的巴拿马血清型肠球菌。
Pub Date : 2020-06-29 eCollection Date: 2020-01-01 DOI: 10.7150/jgen.48656
Anne Busch, Helmut Hotzel, Ulrich Methner

We describe a rare isolate of Salmonella enterica subsp. enterica serovar Panama with an extended-spectrum β-lactamase (ESBL) profile from a German cattle-fattening farm. Applying two next-generation sequencing methods we generated sequences of the genome as well as the plasmids; assembled the draft genome sequence of Salmonella enterica subsp. enterica serovar Panama isolate 18PM0209. Antimicrobial resistance genes, virulence-associated genes and plasmids were analyzed using bioinformatics. Occurrence of multidrug-resistant Salmonella serovars at cattle-fattening farms indicate the need of enhanced surveillance to prevent further spread of these organisms.

我们描述了一种罕见的分离的肠沙门氏菌亚种。具有广谱β-内酰胺酶(ESBL)谱的巴拿马肠菌血清型牛来自德国养牛场。采用两种新一代测序方法,我们获得了基因组序列和质粒;组装肠沙门氏菌亚种基因组序列草图。巴拿马肠血清型分离株18PM0209。采用生物信息学方法对耐药基因、毒力相关基因和质粒进行分析。在养牛场发生的耐多药沙门氏菌血清型表明需要加强监测,以防止这些生物的进一步传播。
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引用次数: 1
crispRdesignR: A Versatile Guide RNA Design Package in R for CRISPR/Cas9 Applications. crispRdesignR:用于CRISPR/Cas9应用的多功能指导RNA设计包。
Pub Date : 2020-05-18 eCollection Date: 2020-01-01 DOI: 10.7150/jgen.41196
Dylan Beeber, Frédéric Jj Chain

The success of CRISPR/Cas9 gene editing applications relies on the efficiency of the single guide RNA (sgRNA) used in conjunction with the Cas9 protein. Current sgRNA design software vary in the details they provide on sgRNA sequence efficiency and usually limit organism choice to a list of developer-selected species. The crispRdesignR package aims to address these limitations by providing comprehensive sequence features of the generated sgRNAs in a single program, which allows users to predict sgRNA efficiency and design sgRNA sequences for systems that currently do not have optimized efficiency scoring methods. crispRdesignR reports extensive information on all designed sgRNA sequences with robust off-target calling and annotation and can be run in a user-friendly graphical interface. The crispRdesignR package is implemented in R and has fully editable code for specialized purposes including sgRNA design in user-provided genomes. The package is platform independent and extendable, with its source code and documentation freely available at https://github.com/dylanbeeber/crispRdesignR.

CRISPR/Cas9基因编辑应用的成功依赖于与Cas9蛋白结合使用的单导RNA (sgRNA)的效率。目前的sgRNA设计软件在提供sgRNA序列效率的细节上各不相同,并且通常将生物选择限制在发育者选择的物种列表中。crispRdesignR软件包旨在通过在单个程序中提供生成的sgRNA的综合序列特征来解决这些限制,使用户可以预测sgRNA的效率,并为目前没有优化效率评分方法的系统设计sgRNA序列。crispRdesignR报告所有设计的sgRNA序列的广泛信息,具有强大的脱靶调用和注释,可以在用户友好的图形界面中运行。crispRdesignR包是用R语言实现的,并且具有完全可编辑的代码,用于特定目的,包括在用户提供的基因组中设计sgRNA。该软件包是平台独立的,可扩展的,其源代码和文档可在https://github.com/dylanbeeber/crispRdesignR免费获得。
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引用次数: 2
Draft genome sequence of Actinomadura sp. K4S16 and elucidation of the nonthmicin biosynthetic pathway. 放线菌 K4S16 基因组序列草案及非桔梗霉素生物合成途径的阐明。
Pub Date : 2020-05-17 eCollection Date: 2020-01-01 DOI: 10.7150/jgen.44650
Hisayuki Komaki, Enjuro Harunari, Natsuko Ichikawa, Akira Hosoyama, Moriyuki Hamada, Kannika Duangmal, Arinthip Thamchaipenet, Yasuhiro Igarashi

Actinomadura sp. K4S16 (=NBRC 110471) is a producer of a novel tetronate polyether compound nonthmicin. Here, we report the draft genome sequence of this strain together with features of the organism and assembly, annotation and analysis of the genome sequence. The 9.6 Mb genome of Actinomadura sp. K4S16 encoded 9,004 putative ORFs, of which 7,701 were assigned with COG categories. The genome contained four type-I polyketide synthase (PKS) gene clusters, two type-II PKS gene clusters, and three nonribosomal peptide synthetase (NRPS) gene clusters. Among the type-I PKS gene (t1pks) clusters, a large t1pks cluster was annotated to be responsible for nonthmicin synthesis based on bioinformatic analyses. We also performed feeding experiments using labeled precursors and propose the biosynthetic pathway of nonthmicin.

Actinomadura sp. K4S16(=NBRC 110471)是一种新型四价聚醚化合物壬苯霉素的生产者。在此,我们报告了该菌株的基因组序列草案、生物特征以及基因组序列的组装、注释和分析。放线菌 K4S16 的 9.6 Mb 基因组编码了 9,004 个推测 ORF,其中 7,701 个被归入 COG 类别。该基因组包含 4 个 I 型多酮合成酶(PKS)基因簇、2 个 II 型 PKS 基因簇和 3 个非核糖体肽合成酶(NRPS)基因簇。在 I 型 PKS 基因簇(t1pks)中,根据生物信息学分析,一个大的 t1pks 基因簇被注释为负责壬苯霉素的合成。我们还利用标记的前体进行了喂养实验,并提出了壬苯霉素的生物合成途径。
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引用次数: 0
Complete Genome Sequence of Leptospira interrogans Strains FMAS_KW1, FMAS_KW2 and FMAS_AW1 Isolated from Leptospirosis Patients from Karawanalla and Awissawella, Sri Lanka. 斯里兰卡Karawanalla和Awissawella钩端螺旋体患者分离株FMAS_KW1、FMAS_KW2和FMAS_AW1的全基因组序列
Pub Date : 2020-04-22 eCollection Date: 2020-01-01 DOI: 10.7150/jgen.43953
Indika Senevirathna, Dinesha Jayasundara, Joshua P Lefler, Kira L Chaiboonm, Janith Warnasekara, Suneth Agampodi, Michael A Matthias, Joseph M Vinetz

Leptospirosis is an important cause of acute undifferentiated fever and complex multisystem febrile diseases in the tropics and subtropics. Understanding the evolution of Leptospira especially as related to the clinical pathogenesis of leptospirosis is facilitated by systematic comparative genomic analysis of human-infecting isolates. Here, we announce the complete genome sequences of three Leptospira strains that were isolated from blood of humans with undifferentiated fever in Sri Lanka.

钩端螺旋体病是热带和亚热带地区急性未分化热和复杂多系统发热疾病的重要病因。了解钩端螺旋体的进化,特别是与钩端螺旋体病的临床发病机制有关,可以通过对人类感染分离株的系统比较基因组分析来促进。在此,我们宣布从斯里兰卡未分化发热患者的血液中分离出的三株钩端螺旋体菌株的完整基因组序列。
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引用次数: 8
A detailed characteristics of bias associated with long runs of homozygosity identification based on medium density SNP microarrays. 基于中密度SNP微阵列的长期纯合性鉴定的详细特征。
Pub Date : 2020-04-07 eCollection Date: 2020-01-01 DOI: 10.7150/jgen.39147
Tomasz Szmatoła, Artur Gurgul, Igor Jasielczuk, Weiwei Fu, Katarzyna Ropka-Molik

In the present study, runs of homozygosity (ROH) detected with the use of a standard bovine 54k single nucleotide polymorphism (SNP) genotyping assay and two different ROH detection approaches, based on 50 (M1) or 15 (M2) consecutive SNPs, were compared with results of whole genome sequencing. Both microarray-based methods accurately recognised medium-sized ROH, however, it was found that M2 method seemed to better than M1 identify short ROH, but highly overestimated their number, leading to numerous false positive calls. Moreover, long ROH identified with microarray data tended to break into shorter segments in sequencing data because of the presence of regions with high heterozygosity within the ROH sequences. This may indicate, that these long ROH are formed by closely positioned shorter homozygous segments that may be of older origin or may be created by two similar but not identical haplotypes, showing minor internal recombination signs. Such finding also suggests that at least some of the results of previous studies in regard to long ROH may be biased leading to inaccurate estimations of genomes autozygosity via ROH classification into length categories.

在本研究中,使用标准牛54k单核苷酸多态性(SNP)基因分型法和基于50 (M1)或15 (M2)个连续SNP的两种不同的ROH检测方法检测纯合子(ROH),并与全基因组测序结果进行了比较。两种基于微阵列的方法都能准确识别中等大小的ROH,然而,我们发现M2方法似乎比M1方法更好地识别短型ROH,但高度高估了它们的数量,导致大量误报。此外,用微阵列数据鉴定的较长的ROH在测序数据中往往会断裂成较短的片段,因为在ROH序列中存在高杂合性的区域。这可能表明,这些长ROH是由位置紧密的较短的纯合片段形成的,这些纯合片段可能来自更古老的起源,或者可能是由两个相似但不相同的单倍型产生的,表现出较小的内部重组迹象。这一发现还表明,至少一些先前关于长ROH的研究结果可能存在偏差,导致通过将ROH分类为长度类别来准确估计基因组自合性。
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引用次数: 3
Draft Genome Sequences of Bifidobacterium animalis Consecutively Isolated from Healthy Japanese Individuals. 日本健康个体连续分离动物双歧杆菌基因组序列草图。
Pub Date : 2020-04-06 eCollection Date: 2020-01-01 DOI: 10.7150/jgen.38516
Tomoya Tsukimi, Tsubasa Watabe, Kazuki Tanaka, Mitsuhiko P Sato, Haruo Suzuki, Masaru Tomita, Shinji Fukuda

Bifidobacterium species are well recognized as probiotics and colonized in various parts of the human body. Here, we report the draft genome sequences of Bifidobacterium animalis isolated from two healthy Japanese volunteers, one of which was sampled twice before and after a 10-year interval. A core genome phylogeny analysis indicated that the strains isolated from the same volunteer were closely related. This paper is the first report of multiple draft genome sequences of B. animalis independently isolated from the same individual and provides insight into the probiotic potential of a member of this species.

双歧杆菌是公认的益生菌,在人体的各个部位都有定植。在这里,我们报告了从两名健康的日本志愿者身上分离的动物双歧杆菌的基因组序列草图,其中一人在10年前和10年后分别取样两次。核心基因组系统发育分析表明,从同一志愿者分离的菌株亲缘关系密切。本文首次报道了从同一个体独立分离的动物双歧杆菌的多个草图基因组序列,并对该物种成员的益生菌潜力进行了深入研究。
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引用次数: 0
Programmable CRISPR interference for gene silencing using Cas13a in mosquitoes. 利用Cas13a对蚊子进行基因沉默的可编程CRISPR干扰。
Pub Date : 2020-03-01 eCollection Date: 2020-01-01 DOI: 10.7150/jgen.43928
Aditi Kulkarni, Wanqin Yu, Alex S Moon, Ashmita Pandey, Kathryn A Hanley, Jiannong Xu

In the CRISPR-Cas systems, Cas13a is an RNA-guided RNA nuclease specifically targeting single strand RNA. We developed a Cas13a mediated CRISPR interference tool to target mRNA for gene silencing in mosquitoes. A Cas13a expressing plasmid was delivered to mosquitoes by intrathoracic injection, and Cas13a transcripts were detectable at least 10 days post-delivery. The target specific crRNA was synthesized in vitro using T7 RNA polymerase. The Cas13a plasmid and target crRNA can be delivered by intrathoracic injection together, or the Cas13a construct can be provided first, and then target crRNA can be given later when appropriate. The machinery was tested in two mosquito species. In Anopheles gambiae, vitellogenin gene was silenced by Cas13a/Vg-crRNA, which was accompanied by a significant reduction in egg production. In Aedes aegypti, the α- and δ-subunits of COPI genes were silenced by Cas13a/crRNA, which resulted in mortality and fragile midguts, reproducing a phenotype reported previously. Co-silencing genes simultaneously is achievable when a cocktail of target crRNAs is given. No detectable collateral cleavages of non-target transcripts were observed in the study. In addition to dsRNA or siRNA mediated RNA interference, the programmable CRISPR interference method offers an alternative to knock down genes in mosquitoes.

在CRISPR-Cas系统中,Cas13a是一种特异性靶向单链RNA的RNA引导RNA核酸酶。我们开发了一种Cas13a介导的CRISPR干扰工具,以mRNA为靶标,对蚊子进行基因沉默。通过胸内注射将Cas13a表达质粒传递给蚊子,在分娩后至少10天可检测到Cas13a转录本。利用T7 RNA聚合酶体外合成目标特异性crRNA。Cas13a质粒和靶crRNA可以胸腔内注射一起给药,也可以先提供Cas13a构建体,合适时再给靶crRNA。这种机器在两种蚊子身上进行了测试。在冈比亚按蚊中,卵黄原蛋白基因被Cas13a/Vg-crRNA沉默,这伴随着产卵量的显著减少。在埃及伊蚊中,COPI基因的α-和δ-亚基被Cas13a/crRNA沉默,导致死亡和脆弱的肠道,再现了先前报道的表型。当给予目标crrna的混合物时,可以同时实现基因共沉默。在研究中未观察到非靶转录本的可检测的侧支切割。除了dsRNA或siRNA介导的RNA干扰外,可编程CRISPR干扰方法还提供了一种敲除蚊子基因的替代方法。
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引用次数: 3
期刊
Journal of Genomics
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