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Draft Genomes Sequences of 11 Geodermatophilaceae Strains Isolated from Building Stones from New England and Indian Stone Ruins found at historic sites in Tamil Nadu, India. 从新英格兰的建筑石块和印度泰米尔纳德邦历史遗址中发现的印度石遗址中分离出的 11 个嗜地肤菌株的基因组序列草案。
Pub Date : 2022-09-21 eCollection Date: 2022-01-01 DOI: 10.7150/jgen.76121
Nathaniel J Ennis, Dhanasekaran Dharumadurai, Joseph L Sevigny, Ryan Wilmot, Sulaiman M Alnaimat, Julia G Bryce, W Kelley Thomas, Louis S Tisa

Metagenomic analysis of stone microbiome from samples collected in New England, USA and Tamil Nadu, India identified numerous Actinobacteria including Geodermatphilaceae. A culture-dependent approach was performed as a companion study with this culture-independent metagenomic analysis of these stone samples and resulted in the isolation of eleven Geodermatphilaceae strains (2 Geodermatophilus and 9 Blastococcus strains). The genomes of the 11 Geodermatphilaceae strains were sequenced and analyzed. The genomes for the two Geodermatophilus isolates, DF1-2 and TF2-6, were 4.45 and 4.75 Mb, respectively, while the Blastococcus genomes ranged in size from 3.98 to 5.48 Mb. Phylogenetic analysis, digital DNA:DNA hybridization (dDDH), and comparisons of the average nucleotide identities (ANI) suggest the isolates represent novel Geodermatophilus and Blastococcus species. Functional analysis of the Geodermatphilaceae genomes provides insight on the stone microbiome niche.

对在美国新英格兰和印度泰米尔纳德邦采集的石材微生物组进行元基因组分析,发现了大量放线菌,包括嗜皮地肤菌科(Geodermatphilaceae)。在对这些石材样本进行不依赖培养的元基因组分析的同时,还进行了依赖培养的方法研究,结果分离出 11 株嗜地肤菌(2 株嗜地肤菌和 9 株高弹性球菌)。对这 11 株地肤菌的基因组进行了测序和分析。两株地肤嗜血杆菌(DF1-2 和 TF2-6)的基因组大小分别为 4.45 和 4.75 Mb,而大肠球菌的基因组大小从 3.98 到 5.48 Mb 不等。系统发育分析、数字 DNA:DNA 杂交(dDDH)和平均核苷酸相同度(ANI)比较表明,这些分离物代表了新的嗜地皮球菌和布拉氏球菌物种。对Geodermatphilaceae基因组的功能分析有助于深入了解石材微生物群的生态位。
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引用次数: 0
Draft Genomes of Symbiotic Frankia Strains AgB32 and AgKG'84/4 from Root Nodules of Alnus Glutinosa growing under Contrasted Environmental Conditions. 对比环境条件下生长的桤木根瘤共生菌株AgB32和AgKG'84/4的基因组绘制
Pub Date : 2022-08-08 eCollection Date: 2022-01-01 DOI: 10.7150/jgen.75779
Philippe Normand, Petar Pujic, Danis Abrouk, Spandana Vemulapally, Trina Guerra, Camila Carlos-Shanley, Dittmar Hahn

The genomes of two nitrogen-fixing Frankia strains, AgB32 and AgKG'84/4, were isolated from spore-containing (spore+) and spore-free (spore-) root nodules of Alnus glutinosa, but they did not sporulate upon reinfection. The two strains are described as representatives of two novel candidate species. Phylogenomic and ANI analyses indicate that each strain represents a novel species within cluster 1, with genome sizes of 6.3 and 6.7 Mb smaller than or similar to those of other cultivated Alnus-infective cluster 1 strains. Genes essential for nitrogen-fixation, clusters of orthologous genes, secondary metabolite clusters and transcriptional regulators analyzed by comparative genomic analyses were typical of those from Alnus-infective cluster 1 cultivated strains in both genomes. Compared to other cultivated Alnus-infective strains with large genomes, those of AgB32 and AgKG'84/4 had lost 380 or 409 genes, among which one hup cluster, one shc gene and the gvp cluster, which indicates genome erosion is taking place in these two strains.

从含孢子(孢子+)和不含孢子(孢子-)的桤木根瘤中分离到两株固氮Frankia菌株AgB32和AgKG'84/4的基因组,但它们在再感染后不产孢。这两个菌株被描述为两个新的候选物种的代表。系统基因组学和ANI分析表明,每一株菌株在集群1中代表一个新种,其基因组大小分别为6.3和6.7 Mb,比其他培养的桤木感染集群1菌株的基因组大小小或相似。固氮必需基因、同源基因簇、次生代谢物簇和转录调节因子在两种基因组中均具有代表性。与其他大基因组培养菌株相比,AgB32和AgKG'84/4分别丢失了380个和409个基因,其中hup基因簇、shc基因簇和gvp基因簇各1个,说明这两个菌株的基因组发生了侵蚀。
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引用次数: 3
Draft Genomes of Six Wild Poisonous Mushrooms. 六种野生毒蘑菇的基因组草图。
Pub Date : 2022-08-01 eCollection Date: 2022-01-01 DOI: 10.7150/jgen.75652
Sittiporn Parnmen, Nattakarn Nooron, Sujitra Sikaphan, Chutimon Uttawichai, Dutsadee Polputpisatkul, Sriprapa Phatsarapongkul, Rungsaeng Chankunasuka, Unchalee Nitma, Chidkamon Thunkhamrak, Nisakorn Palakul, Khwanruan Naksuwankul, Onanong Pringsulaka, Achariya Rangsiruji

Foodborne illnesses caused by wild mushroom poisoning occur globally and have led to food safety concerns. Here, we reported de novo genome assemblies of the six most commonly encountered toxic mushrooms in Thailand. These comprised Amanita brunneitoxicaria, Cantharocybe virosa, Chlorophyllum molybdites, Entoloma mastoideum, Pseudosperma sp. and Russula subnigricans. The nuclear genome sizes of these species ranged from 40 to 77 Mb, with the number of predicted genes ranging from 5,375 to 14,099. The mitogenome sizes varied from 41,555 to 78,907 bp. The resulting draft genomes of these poisonous mushrooms provide insights into toxin-related genes that may be used to establish genetic markers for monitoring mushroom poisoning outbreaks.

野生蘑菇中毒引起的食源性疾病在全球范围内发生,并引起了食品安全问题。在这里,我们报道了泰国六种最常见的有毒蘑菇的从头基因组组装。其中包括褐毒伞菌、斑蝥藻病毒、钼绿藻、乳突肠虫、伪精子和黑穗病菌。这些物种的核基因组大小在40 ~ 77 Mb之间,预测基因数量在5,375 ~ 14,099之间。有丝分裂基因组大小从41,555到78,907 bp不等。由此产生的这些有毒蘑菇的基因组草图提供了对毒素相关基因的见解,可用于建立监测蘑菇中毒爆发的遗传标记。
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引用次数: 1
Draft Genomes of Nitrogen-fixing Frankia Strains Ag45/Mut15 and AgPM24 Isolated from Root Nodules of Alnus Glutinosa 固氮菌株Ag45/Mut15和AgPM24的基因组图谱
Pub Date : 2022-06-06 DOI: 10.7150/jgen.74788
P. Normand, P. Pujić, Danis Abrouk, Spandana Vemulapally, Trina M Guerra, Camila Carlos-Shanley, D. Hahn
The genomes of two nitrogen-fixing Frankia strains, Ag45/Mut15 and AgPM24, isolated from root nodules of Alnus glutinosa are described as representatives of a novel candidate species. Phylogenomic and ANI analyses confirmed that both strains are related to cluster 1 frankiae, and that both strains belong to a novel species. At 6.4 - 6.7 Mb, their genomes were smaller than those of other cultivated Alnus-infective cluster 1 strains but larger than that of the non-cultivated Alnus-infective cluster 1 Sp+ strain AgTrS that was their closest neighbor as assessed by ANI. Comparative genomic analyses identified genes essential for nitrogen-fixation, gene composition as regards COGs, secondary metabolites clusters and transcriptional regulators typical of those from Alnus-infective cluster 1 cultivated strains in both genomes. There were 459 genes present in other cultivated Alnus-infective strains lost in the two genomes, spread over the whole of the genome, which indicates genome erosion is taking place in these two strains.
从Alnus glutinosa根瘤中分离的两个固氮Frankia菌株Ag45/Mut15和AgPM24的基因组被描述为一个新的候选物种的代表。系统基因组学和ANI分析证实,这两株菌株都与第1簇frankiae有关,并且都属于一个新种。在6.4 ~ 6.7 Mb,它们的基因组小于其他栽培的桤木感染簇1菌株,但大于非栽培的桤木感染簇1 Sp+菌株AgTrS,后者是它们最近的邻居。比较基因组分析确定了两个基因组中对固氮、COGs基因组成、次生代谢物簇和转录调节因子所必需的基因,这些基因是桤木感染簇1培养菌株的典型基因。在其他培养的桤木侵染菌株中,有459个基因在这两个基因组中丢失,分布在整个基因组中,这表明这两个菌株的基因组正在发生侵蚀。
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引用次数: 4
Identification of the c.829_832delAATA Deletion Variants in the BRCA1 Gene Associated with Hereditary Breast/Ovarian Cancer ˗ Case Report 与遗传性乳腺癌/卵巢癌相关的BRCA1基因c.829_832delAATA缺失变异体的鉴定——病例报告
Pub Date : 2022-02-14 DOI: 10.7150/jgen.68220
M. Ostrowska, Karolina Olszewska-Bożek, J. Podlodowska, J. Sierocińska-Sawa, J. Wojcierowski
Determination of the BRCA1/BRCA2 mutation status in patients with breast and/or ovarian cancer is commonly performed using various molecular techniques. The use of targeted PCR-based tests only may not be sufficient, as not all possible variants are investigated. In the present study, we used next-generation sequencing (NGS) techniques to identify novel pathogenic variants in BRCA1 and BRCA2. In this study, material (blood and FFPE) collected from a 67-year-old patient with ovarian cancer was used. The presence of hereditary mutations characteristic for the Polish population was examined using Sanger sequencing. BRCA1 and BRCA2 gene exons were amplified using the Devyser BRCA kit and sequenced on the Miniseq. No germline mutations characteristic for the Polish population were detected. However, 12 single nucleotide variants and 2 indels were identified. We found a new deleterious mutation of gene BRCA1 (c.829_832delAATA). To our knowledge, this mutation has not been reported yet in the Polish population and elsewhere. The use of the NGS technique increases the possibility of detecting mutational changes in patients with ovarian and/or breast cancer. Quick determination of pathogenic variants is important to facilitate specific therapy, in addition to the identification of familial predisposition to cancer.
乳腺癌和/或卵巢癌患者BRCA1/BRCA2突变状态的测定通常使用各种分子技术进行。仅仅使用基于pcr的靶向检测可能是不够的,因为并非对所有可能的变异都进行了调查。在本研究中,我们使用下一代测序(NGS)技术来鉴定BRCA1和BRCA2的新致病变异。在这项研究中,材料(血液和FFPE)收集自一位67岁的卵巢癌患者。使用桑格测序检测波兰人群的遗传突变特征。使用Devyser BRCA试剂盒扩增BRCA1和BRCA2基因外显子,并在Miniseq上测序。未发现波兰人群特有的种系突变。然而,鉴定出12个单核苷酸变异和2个索引。我们发现了BRCA1基因的一个新的有害突变(c.829_832delAATA)。据我们所知,这种突变尚未在波兰和其他地方的人群中报道。NGS技术的使用增加了检测卵巢癌和/或乳腺癌患者突变变化的可能性。除了确定癌症的家族易感性外,快速确定致病变异对促进特异性治疗也很重要。
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引用次数: 0
Complete Genome Sequence of Four Strains of Leptospira borgpetersenii serovar Hardjo isolated from Cattle in the Central United States 从美国中部牛身上分离的四株博格彼得森钩端螺旋体血清型Hardjo的全基因组序列
Pub Date : 2022-02-14 DOI: 10.7150/jgen.69822
E. Putz, D. Bayles, D. Alt, J. Nally
Pathogenic species of Leptospira cause leptospirosis, a global zoonotic disease affecting humans and all major livestock species. Cattle act as a reservoir host for L. borgpetersenii serovar Hardjo which colonize the kidneys and reproductive tract from which they are excreted and transmitted to other cattle via urine, semen or uterine discharges. Bovine leptospirosis results in reproductive failure, abortion, stillbirth and loss of milk production, and is an occupational risk for those working with infected animals. A recent study determined that 7.2% of cattle from an abattoir in the central United States were actively shedding pathogenic Leptospira. Here, we report and compare the complete genome sequence of four recent isolates of L. borgpetersenii serovar Hardjo designated strain TC112, TC147, TC129, and TC273.
钩端螺旋体病是一种影响人类和所有主要牲畜的全球性人畜共患疾病。牛是L.borgpetersenii serovar Hardjo的宿主,其定植于肾脏和生殖道,通过尿液、精液或子宫分泌物排出并传播给其他牛。牛钩端螺旋体病会导致繁殖失败、流产、死产和产奶量减少,对感染动物的工作人员来说是一种职业风险。最近的一项研究确定,美国中部屠宰场7.2%的牛正在积极脱落致病性钩端螺旋体。在这里,我们报道并比较了博尔格彼得森乳杆菌血清型Hardjo指定菌株TC112、TC147、TC129和TC273的四个最新分离株的全基因组序列。
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引用次数: 3
GTQC: Automated Genotyping Array Quality Control and Report. GTQC:自动基因分型阵列质量控制和报告。
Pub Date : 2022-02-14 eCollection Date: 2022-01-01 DOI: 10.7150/jgen.69860
Shilin Zhao, Limin Jiang, Hui Yu, Yan Guo

Genotyping array is the most economical approach for conducting large-scale genome-wide genetic association studies. Thorough quality control is key to generating high integrity genotyping data and robust results. Quality control of genotyping array is generally a complicated process, as it requires intensive manual labor in implementing the established protocols and curating a comprehensive quality report. There is an urgent need to reduce manual intervention via an automated quality control process. Based on previously established protocols and strategies, we developed an R package GTQC (GenoTyping Quality Control) to automate a majority of the quality control steps for general array genotyping data. GTQC covers a comprehensive spectrum of genotype data quality metrics and produces a detailed HTML report comprising tables and figures. Here, we describe the concepts underpinning GTQC and demonstrate its effectiveness using a real genotyping dataset. R package GTQC streamlines a majority of the quality control steps and produces a detailed HTML report on a plethora of quality control metrics, thus enabling a swift and rigorous data quality inspection prior to downstream GWAS and related analyses. By significantly cutting down on the time on genotyping quality control procedures, GTQC ensures maximum utilization of available resources and minimizes waste and inefficient allocation of manual efforts. GTQC tool can be accessed at https://github.com/slzhao/GTQC.

基因分型阵列是进行大规模全基因组遗传关联研究最经济的方法。彻底的质量控制是生成高完整性基因分型数据和可靠结果的关键。基因分型阵列的质量控制通常是一个复杂的过程,因为它需要大量的人工来执行既定的方案和编制全面的质量报告。目前迫切需要通过自动化质量控制流程来减少人工干预。基于之前建立的协议和策略,我们开发了一个 R 软件包 GTQC(基因分型质量控制),以自动完成一般阵列基因分型数据的大部分质量控制步骤。GTQC 涵盖了基因型数据质量指标的全面范围,并生成一份包含表格和图表的详细 HTML 报告。在此,我们将介绍 GTQC 的基本概念,并使用一个真实的基因分型数据集演示其有效性。R软件包GTQC简化了大部分质量控制步骤,并能生成关于大量质量控制指标的详细HTML报告,因此能在下游GWAS和相关分析之前快速、严格地检查数据质量。通过大幅缩短基因分型质量控制程序的时间,GTQC 可确保最大限度地利用现有资源,最大限度地减少浪费和低效的人工分配。GTQC工具可在https://github.com/slzhao/GTQC。
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引用次数: 0
Identification and Characterization of a Potential Antibiotic Producing Strain of Pantoea ananatis. 一种潜在抗生素生产菌株的鉴定和特征描述。
Pub Date : 2022-02-07 eCollection Date: 2022-01-01 DOI: 10.7150/jgen.70066
Michael J LaGier, Mark McDaniel, Alyssa Ragner, Amber Castillo

Antibiotic resistance continues to be a significant public health challenge. Soil bacteria represent a potential source of yet to be discovered antimicrobials. The screening of Iowa (United States) soils yielded the identification of a strain of Pantoea ananatis (MMB-1), which displayed an antimicrobial-producing phenotype against a bacterium (Bacillus subtilis) representative of Gram-positive bacteria. Crude, organic, extracts of MMB-1 retained the anti-microbial activity. The draft genome of strain MMB-1 contains a total of 4,634,340 bp, and 4,624 protein-encoding genes. Consistent with phenotypic observation, the genome of MMB-1 encodes for a number of putative secondary metabolite biosynthetic gene clusters, including those known to be involved in the production of the antibiotics lankacidin C and bottromycin. This study increases our overall understanding of Panteoa as a group, and is also consistent with the notion that members of this genus have significant potential as useful natural product producers.

抗生素耐药性仍然是一个重大的公共卫生挑战。土壤细菌是尚未发现的抗菌素的潜在来源。通过对美国爱荷华州土壤的筛选,发现了一株泛函菌(MMB-1),它对革兰氏阳性细菌的代表细菌(枯草芽孢杆菌)具有抗菌表型。MMB-1 的有机粗提取物保留了抗微生物活性。菌株 MMB-1 的基因组草案共包含 4,634,340 bp 和 4,624 个编码蛋白质的基因。与表型观察结果一致的是,MMB-1 的基因组编码了许多推测的次级代谢物生物合成基因簇,包括那些已知参与生产抗生素兰卡霉素 C 和瓶霉素的基因簇。这项研究增加了我们对 Panteoa 作为一个类群的总体了解,也与该属成员作为有用的天然产品生产者具有巨大潜力的观点相一致。
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引用次数: 0
Circ-0036602 Acts As a Sponge of MiR-34a-5p and MiR-431-5p to Promote Cervical Cancer Proliferation and Invasion. Circ-0036602作为MiR-34a-5p和MiR-431-5p的海绵促进宫颈癌的增殖和侵袭
Pub Date : 2022-01-11 eCollection Date: 2022-01-01 DOI: 10.7150/jgen.62458
Xia Ye, Biqing Zhu, Jingjing Han, Jian Huang, Yaqin Wu

Background: Cervical cancer (CC) is one of the most common female malignancies worldwide. An increasing body of evidence suggests that circular RNAs (circRNAs) participate in the pathogenesis of various cancers, including CC. However, the expression profile and underlying molecular mechanisms remain largely unknown. Methods: In this study, high throughput sequencing was applied to identify circRNA in HPV-16 positive CC tissues. Quantitative real-time PCR (qRT-PCR) was performed to validate the expression in CC tissues and cell lines. RNase R treatment, gel-electrophoresis, and RNA fluorescent in situ hybridization (FISH) were used to characterize the circRNAs. Subsequently, the Cell Counting Kit-8 assay (CCK8), transwell and wound healing assays were performed to assess circRNA function. Meanwhile, dual-luciferase reporter and western blot were used to clarify the associated molecular mechanisms. Results: Circ0036602 was upregulated in HPV-16 positive CC and correlated with a poor prognosis. Moreover, circ0036602 expression significantly correlated with the clinicopathologic characteristics. Knockdown of circ0036602 inhibited CC cell proliferation, migration, and invasion. Further studies showed that circ0036602 could bind to miR-34-5p and miR-431-5p to regulate the expression of the target gene HMGB1. Conclusions: Taken together, our findings suggest that circ0036602 is a tumor-promoting circRNA that promotes CC cells by sponging miR-34-5p and miR-431-5p to regulate HMGB1. Circ0036602 has huge prospects as a potential therapeutic target for CC patients.

背景:宫颈癌是世界范围内最常见的女性恶性肿瘤之一。越来越多的证据表明,环状rna (circRNAs)参与了包括CC在内的多种癌症的发病机制,然而,其表达谱和潜在的分子机制在很大程度上仍然未知。方法:在本研究中,采用高通量测序技术鉴定HPV-16阳性CC组织中的circRNA。采用实时荧光定量PCR (qRT-PCR)验证其在CC组织和细胞系中的表达。使用RNase R处理、凝胶电泳和RNA荧光原位杂交(FISH)来表征环状RNA。随后,进行细胞计数试剂盒-8试验(CCK8)、transwell和伤口愈合试验来评估circRNA的功能。同时,利用双荧光素酶报告基因和western blot技术阐明了相关的分子机制。结果:Circ0036602在HPV-16阳性CC中表达上调,并与不良预后相关。此外,circ0036602的表达与临床病理特征显著相关。敲低circ0036602可抑制CC细胞增殖、迁移和侵袭。进一步研究表明circ0036602可结合miR-34-5p和miR-431-5p调控靶基因HMGB1的表达。综上所述,我们的研究结果表明circ0036602是一种促肿瘤的circRNA,通过海绵miR-34-5p和miR-431-5p调节HMGB1来促进CC细胞。Circ0036602作为CC患者的潜在治疗靶点具有巨大的前景。
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引用次数: 4
Comparative analysis of mRNA transcripts of HT-29 cell line expressed in identical quantities for pathogenic E. coli strains UM146 and UM147 with control Escherichia coli Nissle 1917. 致病性大肠杆菌UM146和UM147与对照大肠杆菌Nissle 1917表达量相同的HT-29细胞系mRNA转录物的比较分析
Pub Date : 2022-01-01 DOI: 10.7150/jgen.67277
Roman Kotłowski

Aim of study was comparative analysis of mRNA transcripts of HT-29 cell line, expressed in identical quantities for the combination of pathogenic and non-pathogenic Escherichia coli strains. HT-29 confluent monolayers infection with two pathogenic E. coli strains UM146 and UM147 resulted in two sets of mRNA transcripts that were identical with RNA transcripts obtained for non-pathogenic one strain E. coli Nissle 1917. In this study genome-wide experiments were conducted using expression microarray-system. Only one common mRNA transcript coding for CCDC65 gene was equally expressed by HT-29 cells after incubation challenge with three different E. coli strains used. This gene and its bacterial analogue are important in the ciliary or flagellar motility, respectively. Altogether, 78 and 81 HT-29 mRNA transcripts for E. coli UM146 and E. coli UM147 had identical RNA quantity in comparison to the response obtained for non-pathogenic E. coli Nissle 1917 interactions with HT-29 monolayers. Specific analysis using REACTOME and agriGO terms enrichment data-mining tools as well as word-cloud analysis allowed for identification the most important processes characteristic during HT-29 cell line infections for each pathogenic E. coli strain used. The importance of results may contribute to recognition of those processes during bacterial infections that are identical with processes arising from human interaction with non-pathogenic strains that belong to the same bacterial species.

研究的目的是比较分析致病性和非致病性大肠杆菌菌株联合表达量相同的HT-29细胞系mRNA转录本。两株致病性大肠杆菌UM146和UM147感染HT-29融合单层膜,得到两组mRNA转录物,与非致病性大肠杆菌Nissle 1917获得的RNA转录物相同。本研究采用表达微阵列系统进行全基因组实验。在三种不同的大肠杆菌孵育后,HT-29细胞只表达了一种编码CCDC65基因的常见mRNA转录物。该基因及其细菌类似物分别在纤毛或鞭毛运动中起重要作用。总的来说,大肠杆菌UM146和大肠杆菌UM147的78和81个HT-29 mRNA转录物与非致病性大肠杆菌Nissle 1917与HT-29单层相互作用的反应相比,具有相同的RNA数量。使用REACTOME和agriGO术语丰富数据挖掘工具以及词云分析进行特异性分析,可以识别所使用的每种致病性大肠杆菌菌株在HT-29细胞系感染期间最重要的过程特征。结果的重要性可能有助于认识细菌感染期间的那些过程,这些过程与人类与属于同一细菌种类的非致病性菌株相互作用所产生的过程相同。
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引用次数: 0
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Journal of Genomics
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