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Aeromonas salmonicida isolates from Canada demonstrate wide distribution and clustering among mesophilic strains. 从加拿大分离的沙门氏菌气单胞菌在嗜酸性菌株中具有广泛的分布和聚集性。
IF 3.1 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2023-05-01 DOI: 10.1139/gen-2022-0086
Sabrina A Attéré, Cynthia Gagné-Thivierge, Valérie E Paquet, Gabrielle R Leduc, Antony T Vincent, Steve J Charette

All the 36 known species to date of the genus Aeromonas are mesophilic except the species Aeromonas salmonicida, which includes both psychrophilic and mesophilic subspecies. For 20 years, more and more mesophilic A. salmonicida strains have been discovered. Only A. salmonicida subsp. pectinolytica has officially been classified as a mesophilic subspecies. Most mesophiles have been isolated in hot countries. We present, for the first time, the characterization of two new mesophilic isolates from Quebec (Canada). Phenotypic and genomic characterizations were carried out on these strains, isolated from dead fish from a fish farm. Isolates 19-K304 and 19-K308 are clearly mesophiles, virulent to the amoeba Dictyostelium discoideum, a surrogate host, and close to strain Y577, isolated in India. To our knowledge, this is the first time that mesophilic strains isolated from different countries are so similar. The major difference between the isolates is the presence of plasmid pY47-3, a cryptic plasmid that sometimes presents in mesophilic strains. More importantly, our extensive phylogenetic analysis reveals two well-defined clades of mesophilic strains with psychrophiles associated with one of these clades. This helps to have a better understanding of the evolution of this species and the apparition of psychrophilic subspecies.

所有已知的36种气单胞菌属都是嗜温的,除了沙门氏菌气单胞菌,它包括嗜温亚种和嗜温亚种。20年来,越来越多的嗜酸性沙门氏菌菌株被发现。只有沙门氏菌亚种。果胶溶菌已被正式归类为嗜中温亚种。大多数嗜温菌已在炎热国家被分离出来。我们首次报道了来自加拿大魁北克省的两种新的中温菌株的特征。对这些从一个养鱼场的死鱼中分离出来的菌株进行了表型和基因组鉴定。分离株19-K304和19-K308显然是嗜中菌,对代用寄主盘状变形虫(Dictyostelium disideum)有毒力,与在印度分离的菌株Y577接近。据我们所知,这是第一次从不同国家分离的嗜温菌株如此相似。分离株之间的主要区别是存在质粒pY47-3,这是一种有时出现在中温菌株中的隐质粒。更重要的是,我们广泛的系统发育分析揭示了两个定义明确的中温菌株分支,其中一个分支与嗜冷菌相关。这有助于更好地理解这个物种的进化和嗜冷亚种的出现。
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引用次数: 1
Characterization of the chloroplast genome of medicinal herb Polygonatum cyrtonema and identification of molecular markers by comparative analysis. 中药黄精叶绿体基因组的鉴定及分子标记的比较分析。
IF 3.1 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2023-04-01 DOI: 10.1139/gen-2022-0087
Zhen-Ni Xie, Jia Lao, Hao Liu, Wan-Xiang Zhang, Wei He, Can Zhong, Jing Xie, Shui-Han Zhang, Jian Jin

Polygonatum cyrtonema Hua is a traditional Chinese herb medicine, and it is widely distributed in China. The intrageneric taxonomy and phylogenetic relationships within Polygonatum have long been controversial due to their morphological similarity and lacking special DNA barcodes. In this paper, the complete chloroplast genome is a relatively conserved quadripartite structure including a large single copy region of 84 711 bp, a small single copy region of 18 210 bp, and a pair of inverted repeats region of 26 142 bp. A total of 342 simple sequence repeats were identified, and most of them were found to be composed of A/T, including 126 mono-nucleotides and 179 di-nucleotides. Nucleotide diversity was analyzed and eight highly variable regions (psbl∼trnT-CGU, atpF∼atpH, trnT-GGU∼psbD, psaJ∼rps20, trnL-UAG∼ndhD, ndhG∼ndhl, ndhA, and rpl32∼ccsA) were identified as potential molecular markers. Phylogenetic analysis based on the whole chloroplast genome showed that P. cyrtonema, within the family Asparagaceae, is closely related to Polygonatum sibiricum and Polygonatum kingianum. The sequence matK, trnT-GGU & ccsA, and ndhG∼ndhA were identified as three DNA barcodes. The assembly and comparative analysis of P. cyrtonema complete chloroplast genome will provide essential molecular information about the evolution and molecular biology for further study.

黄精是一种传统的中草药,在中国广泛分布。黄精属植物的属内分类和系统发育关系由于形态相似性和缺乏特殊的DNA条形码而长期存在争议。在本文中,完整的叶绿体基因组是一个相对保守的四分体结构,包括84 711 bp的大单拷贝区,18 210 bp的小单拷贝区和26 142 bp的一对反向重复区。共鉴定出342个简单重复序列,其中大部分由A/T组成,包括126个单核苷酸和179个双核苷酸。分析了核苷酸多样性,并确定了8个高度可变的区域(psbl ~ trnT-CGU、atpF ~ atpH、trnt ~ ggu ~ psbD、psaJ ~ rps20、trl - uag ~ ndhD、ndhG ~ ndhl、ndhA和rpl32 ~ ccsA)作为潜在的分子标记。基于全叶绿体基因组的系统发育分析表明,天门冬科蓼属植物cyrtonema与黄精(Polygonatum sibiricum)和黄精(Polygonatum kingianum)亲缘关系较近。序列matK、trnT-GGU & ccsA和ndhG ~ ndhA被鉴定为三个DNA条形码。胞浆假体叶绿体全基因组的组装和比较分析,将为胞浆假体进化和分子生物学的进一步研究提供重要的分子信息。
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引用次数: 0
Pig fetal skeletal muscle development is associated with genome-wide DNA hypomethylation and corresponding alterations in transcript and microRNA expression. 猪胎儿骨骼肌发育与全基因组DNA低甲基化以及相应的转录物和microRNA表达改变有关。
IF 3.1 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2023-04-01 DOI: 10.1139/gen-2022-0008
R J Corbett, L M Ford, N E Raney, J M Grabowski, C W Ernst

Fetal myogenesis represents a critical period of porcine skeletal muscle development and requires coordinated expression of thousands of genes. Epigenetic mechanisms, including DNA methylation, drive transcriptional regulation during development; however, these processes are understudied in developing porcine tissues. We performed bisulfite sequencing to assess DNA methylation in pig longissimus dorsi muscle at 41- and 70-days gestation (dg), as well as RNA- and small RNA-sequencing to identify coordinated changes in methylation and expression between myogenic stages. We identified 45 739 differentially methylated regions (DMRs) between stages, and the majority (N = 34 232) were hypomethylated at 70 versus 41 dg. Integration of methylation and transcriptomic data revealed strong associations between differential gene methylation and expression. Differential miRNA methylation was significantly negatively correlated with abundance, and dynamic expression of assayed miRNAs persisted postnatally. Motif analysis revealed significant enrichment of myogenic regulatory factor motifs among hypomethylated regions, suggesting that DNA hypomethylation may function to increase accessibility of muscle-specific transcription factors. We show that developmental DMRs are enriched for GWAS SNPs for muscle- and meat-related traits, demonstrating the potential for epigenetic processes to influence phenotypic diversity. Our results enhance understanding of DNA methylation dynamics of porcine myogenesis and reveal putative cis-regulatory elements governed by epigenetic processes.

胎儿肌发生是猪骨骼肌发育的关键时期,需要数千个基因的协调表达。表观遗传机制,包括DNA甲基化,在发育过程中驱动转录调控;然而,这些过程在猪组织发育中的研究还不够充分。我们对妊娠41天和70天(dg)的猪背最长肌进行亚硫酸盐测序,以评估其DNA甲基化,并对RNA和小RNA进行测序,以确定在肌生成阶段甲基化和表达的协调变化。我们确定了45 739个不同阶段的差异甲基化区域(DMRs),大多数(N = 34 232)在70 dg和41 dg时低甲基化。甲基化和转录组数据的整合揭示了差异基因甲基化和表达之间的强烈关联。差异miRNA甲基化与丰度显着负相关,并且所测miRNA的动态表达在出生后持续存在。基序分析显示,在低甲基化区域中,肌生成调节因子基序显著富集,表明DNA低甲基化可能有助于增加肌肉特异性转录因子的可及性。我们发现,发育中的DMRs富含肌肉和肉类相关性状的GWAS snp,这表明表观遗传过程可能影响表型多样性。我们的研究结果增强了对猪肌肉发生DNA甲基化动力学的理解,并揭示了表观遗传过程中可能的顺式调控元件。
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引用次数: 1
High nucleotide similarity of three Copia lineage LTR retrotransposons among plant genomes. 三个Copia谱系LTR反转录转座子在植物基因组中的高核苷酸相似性。
IF 3.1 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2023-03-01 DOI: 10.1139/gen-2022-0026
Simon Orozco-Arias, Mathilde Dupeyron, David Gutierrez-Duque, Reinel Tabares-Soto, Romain Guyot

Transposable elements (TEs) are mobile elements found in the majority of eukaryotic genomes. TEs deeply impact the structure and evolution of chromosomes and can induce mutations affecting coding genes. In plants, the major group of TEs is long terminal repeat retrotransposons (LTR-RTs). They are classified into superfamilies (Gypsy, Copia) and subclassified into lineages. Horizontal transfer (HT), defined as the nonsexual transmission of genetic material between species, is a process allowing LTR-RTs to invade a new genome. Although this phenomenon was considered rare, recent studies demonstrate numerous transfers of LTR-RTs. This study aims to determine which LTR-RT lineages are shared with high similarity among 69 plant genomes. We identified and classified 88 450 LTR-RTs and determined 143 cases of high similarities between pairs of genomes. Most of them involved three Copia lineages (Oryco/Ivana, Retrofit/Ale, and Tork/Tar/Ikeros). A detailed analysis of three cases of high similarities involving Tork/Tar/Ikeros group shows an uneven distribution in the phylogeny of the elements and incongruence with between phylogenetic trees topologies, indicating they could be originated from HTs. Overall, our results suggest that LTR-RT Copia lineages share outstanding similarity between distant species and may likely be involved in HT mechanisms more frequent than initially estimated.

转座因子(te)是在大多数真核生物基因组中发现的可移动元件。te深刻影响染色体的结构和进化,并能诱导影响编码基因的突变。在植物中,te的主要类群是长末端重复反转录转座子(LTR-RTs)。他们被分为大家族(吉普赛,科皮亚)和分支。水平转移(HT)被定义为物种间遗传物质的非性传播,是一个允许LTR-RTs入侵新基因组的过程。虽然这种现象被认为是罕见的,但最近的研究表明,ltr - rt有许多转移。本研究旨在确定69个植物基因组中哪些LTR-RT谱系具有高相似性。我们鉴定并分类了88450个LTR-RTs,并确定了143个基因组对之间的高相似性。其中大多数涉及三个Copia血统(Oryco/Ivana, Retrofit/Ale和Tork/Tar/Ikeros)。通过对3例涉及Tork/Tar/Ikeros类群的高相似性的详细分析,发现它们在系统发育上分布不均匀,系统发育树拓扑结构不一致,表明它们可能起源于HTs。总的来说,我们的研究结果表明,LTR-RT Copia谱系在远缘物种之间具有突出的相似性,可能比最初估计的更频繁地参与高温机制。
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引用次数: 0
Cytogenetic characterization of solitary wasp Ancistrocerus flavomarginatus (Brèthes, 1906) (Hymenoptera, Vespidae) with insights into the chromosomal evolution in the genus. 黄斑孤蜂(Ancistrocerus flavomarginatus, br<s:1>, 1906)(膜翅目,蜂科)的细胞遗传学特征及其属染色体进化的认识。
IF 3.1 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2023-03-01 DOI: 10.1139/gen-2022-0095
Mara Garcia Tavares, Gisele Amaro Teixeira

Cytogenetic studies have enabled the characterization of the chromosomal macrostructure and microstructure and have contributed to the understanding of the evolution of wasp karyotypes. However, studies on Eumeninae solitary wasps are scarce. In this study, we characterized the karyotype of Ancistrocerus flavomarginatus (Brèthes, 1906) and compared it with previous data from other Ancistrocerus (Wesmael, 1836) species to shed light on the chromosomal diversity of the genus. A chromosome number of 2n = 24 in females and n = 12 in males was observed. Comparing the A. flavomarginatus karyotype with that of another Ancistrocerus species showed variations in the morphology of some chromosomal pairs. The presence of two larger chromosome pairs, almost entirely heterochromatic, and the predominance of subtelocentric chromosomes with heterochromatic short arms in A. flavomarginatus support the occurrence of fissions in Ancistrocerus. A single site of ribosomal genes was observed in A. flavomarginatus, in addition to a size polymorphism of these rDNA clusters between the homologues of some analyzed females. This polymorphism may originate from duplications/deletions due to unequal crossing-over or amplification via transposable elements. The (GA)15 microsatellite is located exclusively in euchromatic regions. Our data show that different rearrangements seem to shape chromosomal evolution in Ancistrocerus species.

细胞遗传学研究使染色体宏观结构和微观结构的表征成为可能,并有助于理解黄蜂核型的进化。然而,对独居胡蜂的研究却很少。本研究对Ancistrocerus flavomarginatus (br thes, 1906)的核型进行了鉴定,并与其他Ancistrocerus (Wesmael, 1836)种的资料进行了比较,以阐明该属的染色体多样性。雌性染色体数为2n = 24,雄性染色体数为n = 12。将黄尾猿核型与另一种蛇尾猿核型进行比较,发现某些染色体对的形态存在差异。A. flavomarginatus中两对较大的几乎完全异色的染色体的存在,以及亚远心短臂异色染色体的优势,支持了Ancistrocerus分裂的发生。黄颡鱼的核糖体基因只有一个位点,而且这些rDNA簇的大小在一些雌性同系物之间存在多态性。这种多态性可能源于不平等交叉或转座元件扩增造成的重复/缺失。(GA)15微卫星仅位于正色区域。我们的数据显示,不同的重排似乎塑造了蛇尾龙物种的染色体进化。
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引用次数: 0
Characterization and molecular evolution analysis of Periploca forrestii inferred from its complete chloroplast genome sequence. 林柏属植物叶绿体全基因组序列的鉴定与分子进化分析。
IF 3.1 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2023-02-01 DOI: 10.1139/gen-2022-0050
Tianze Wu, Gang Deng, Qinggang Yin, Shilin Chen, Yongping Zhang, Bo Wang, Li Xiang, Xia Liu

Periploca forrestii, a medicinal plant of the family Apocynaceae, is known as an effective and widely used clinical prescription for the treatment of rheumatoid diseases. In this study, we de novo sequenced and assembled the completement chloroplast (cp) genome of P. forrestii based on combined Oxford Nanopore PromethION and Illumina data. The cp genome was 153 724 bp in length and had four subregions. Moreover, an 84 433 bp large single-copy and a 17 731 bp small single-copy were separated by 25 780 bp inverted repeats (IRs). The cp genome included 132 genes with 18 duplicates in the IRs. A total of 45 repeat structures and 183 simple sequence repeats were detected. Codon usage showed a bias toward A/T-ending codons. A comparative study of Apocynaceae revealed that an IR expansion occurred on P. forrestii. The Ka/Ks values of eight species of Apocynaceae suggested that positive selection was exerted on the psaI and ycf2 genes, which might reflect specific adaptions to the P. forrestii particular growth environment. Phylogenetic analysis indicated that Periplocoideae was a sister to Asclepiadoideae, forming a monophyletic group in the family Apocynaceae. This study provided an important P. forrestii genomic resource for future evolutionary studies and the phylogenetic reconstruction of the family Apocynaceae.

夹竹桃是夹竹桃科的一种药用植物,是临床上广泛使用的治疗类风湿性疾病的有效方药。在这项研究中,我们基于牛津纳米孔PromethION和Illumina的联合数据重新测序和组装了森林p.r restii的叶绿体(cp)基因组。cp基因组全长153 724 bp,有4个亚区。用25 780 bp的反向重复序列(IRs)分离了84 433 bp的大单拷贝和17 731 bp的小单拷贝。cp基因组包括132个基因,在IRs中有18个重复。共检测到45个重复结构和183个简单序列重复。密码子的使用倾向于a / t结尾的密码子。通过对夹竹桃科植物的比较研究发现,林松发生了IR扩展。8种夹竹桃科植物的Ka/Ks值表明,psaI和ycf2基因发生了正选择,这可能反映了夹竹桃对特定生长环境的特异性适应。系统发育分析表明,该科为夹竹桃科的姐妹科,是夹竹桃科的一个单系类群。该研究为夹竹桃科植物的进化研究和系统发育重建提供了重要的基因组资源。
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引用次数: 0
Differentiation of Lingxiaohua and Yangjinhua by chloroplast genome sequencing and DNA barcoding markers. 利用叶绿体基因组测序和DNA条形码标记进行凌霄花和杨金花的分化。
IF 3.1 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2023-02-01 DOI: 10.1139/gen-2022-0063
Hiu-Lam Ngai, Bobby Lim-Ho Kong, David Tai-Wai Lau, P C Shaw

Lingxiaohua (Campsis Flos, Campsis grandiflora (Thunb.) K. Schum) is a medicinal herb used for promoting diuresis and treating blood-related disorders by the promotion of blood circulation. It also possesses anti-inflammatory and antioxidative properties. This non-poisonous plant is frequently confused with poisonous Yangjinhua (Daturae Metelis Flos, Datura metel Linnaeus) in the market, resulting in serious anticholinergic poisoning. The confusion of these two herbs is due to the similarity in their appearances. In our study, we compared the complete chloroplast genomes of the two plants and found that they are very different in terms of their gene content and gene arrangement. There were also significant differences in the number and repeating motifs of microsatellites and complex repeats. We used universal primers for the amplification of rbcL, matK, psbA-trnH, and ITS2 regions and successfully differentiated the two plants. Furthermore, we designed two pairs of primers based on the nucleotide differences in chloroplast genomes at the rps14 and rpoC1 regions to provide additional authentication markers. The universal primers and specific primers when used together can accurately discriminate Lingxiaohua and Yangjinhua.

凌霄花(山茱萸,桔梗)舒姆(K. Schum)是一种通过促进血液循环来促进利尿和治疗血液相关疾病的草药。它还具有抗炎和抗氧化的特性。这种无毒植物在市场上经常与有毒的杨金花(Daturae Metelis Flos, Datura metel Linnaeus)混淆,导致严重的抗胆碱能中毒。这两种草药的混淆是由于它们的外观相似。在我们的研究中,我们比较了两种植物的完整叶绿体基因组,发现它们在基因含量和基因排列上有很大的不同。微卫星序列和复重复序列在数目和重复基序上也存在显著差异。我们使用通用引物扩增rbcL、matK、psbA-trnH和ITS2区域,成功分化了两株植物。此外,我们根据叶绿体基因组rps14和rpoC1区域的核苷酸差异设计了两对引物,以提供额外的鉴定标记。通用引物和特异引物结合使用,能准确区分凌霄花和阳金花。
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引用次数: 2
Note of appreciation. 表示感谢。
IF 3.1 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2023-01-01 DOI: 10.1139/gen-2022-0104
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引用次数: 0
Complete genome analysis of Pseudomonas furukawaii ZS1 isolated from grass carp (Ctenopharyngodon idellus) culture water. 草鱼养殖水体古卡假单胞菌ZS1的全基因组分析。
IF 3.1 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2023-01-01 DOI: 10.1139/gen-2022-0055
Shuhui Niu, Wangbao Gong, Zhifei Li, Kai Zhang, Guangjun Wang, Ermeng Yu, Yun Xia, Jingjing Tian, Hongyan Li, Jiajia Ni, Jun Xie

Pseudomonas furukawaii ZS1, isolated from grass carp (Ctenopharyngodon idellus) culture water, exhibits efficient aerobic nitrate reduction without nitrite accumulation; however, the molecular pathway underlying this aerobic nitrate reduction remains unclear. In this study, we constructed a complete genome map of P. furukawaii ZS1 and performed a comparative genomic analysis with a reference strain. The results showed that P. furukawaii ZS1 genome was 6 026 050 bp in size and contained 5427 predicted protein-coding sequences. The genome contained all the necessary genes for the dissimilatory nitrate reduction to ammonia pathway but lacked those for the assimilatory nitrate reduction pathway; additionally, genes that convert ammonia to organic nitrogen were also identified. The presence of putative genes associated with the nitrogen and oxidative phosphorylation pathways implied that ZS1 can perform respiration and nitrate reduction simultaneously under aerobic conditions, so that nitrite is rapidly consumed for detoxication by denitrification. The aim of this study is to indicate the great potential of strain ZS1 for future full-scale applications in aquaculture. This work provided insights at the molecular level on the nitrogen metabolic pathways in Pseudomonas species. The understanding of nitrogen metabolic pathways also provides significant molecular information for further Pseudomonas species modification and development.

从草鱼(Ctenopharyngodon idellus)培养水中分离出的古卡假单胞菌ZS1,表现出有效的好氧硝酸盐还原而不积累亚硝酸盐;然而,这种有氧硝酸盐还原的分子途径尚不清楚。在本研究中,我们构建了P. furukawaii ZS1的完整基因组图谱,并与参考菌株进行了比较基因组分析。结果表明,古氏疟原虫ZS1基因组大小为6 ~ 26 ~ 050 bp,包含5427个预测蛋白编码序列。基因组中含有异化硝态氮还原氨途径所需的全部基因,但缺乏同化硝态氮还原途径所需的基因;此外,还发现了将氨转化为有机氮的基因。与氮和氧化磷酸化途径相关的假定基因的存在表明,ZS1在有氧条件下可以同时进行呼吸和硝酸盐还原,从而通过反硝化作用迅速消耗亚硝酸盐进行解毒。本研究的目的是指出菌株ZS1在未来水产养殖中大规模应用的巨大潜力。这项工作在分子水平上对假单胞菌物种的氮代谢途径提供了见解。对氮代谢途径的了解也为假单胞菌的进一步改造和发育提供了重要的分子信息。
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引用次数: 1
NRAS promotes the proliferation of melanocytes to increase melanin deposition in Rex rabbits. NRAS促进黑素细胞增殖,增加獭兔黑色素沉积。
IF 3.1 3区 生物学 Q2 Biochemistry, Genetics and Molecular Biology Pub Date : 2023-01-01 DOI: 10.1139/gen-2021-0111
Shaocheng Bai, Shuaishuai Hu, Yingying Dai, Rongshuai Jin, Chen Zhang, Fan Yao, Qiaoqin Weng, Pin Zhai, Bohao Zhao, Xinsheng Wu, Yang Chen

Melanocytes play a major role in the formation of mammalian fur color and are regulated by several genes. Despite playing the pivotal role in the study of melanoma, the mechanistic role of NRAS (neuroblastoma RAS viral oncogene homolog) in the formation of mammalian epidermal color is still elusive. First of all, the expression levels of NRAS mRNA and protein in the dorsal skin of different colored Rex rabbits were detected by qRT-PCR and Western blot. Then, the subcellular localization of NRAS was identified in melanocytes by indirect immunofluorescence. Next, the expression of NRAS was overexpressed and knocked down in melanocytes, and its efficiency was verified by qRT-PCR and Western blot. Subsequently, NaOH, CCK-8, and Annexin V-FITC were used to verify the changes in melanin content, proliferation, and apoptosis in melanocytes. Finally, we analyzed the regulation of NRAS on other genes (MITF, TYR, DCT, PMEL, and CREB) that affect melanin production. In silico studies showed NRAS as a stable and hydrophilic protein, and it is localized in the cytoplasm and nucleus of melanocytes. The mRNA and protein expression levels of NRAS were significantly different in skin of different colored Rex rabbits, and the highest level was found in black skin (P < 0.01). Moreover, the NRAS demonstrated impact on the proliferation, apoptosis, and melanin production of melanocytes (P < 0.05), and the strong correlation of NRAS with melanin-related genes was evidently observed (P < 0.05). Our results suggested that NRAS can be used as a gene that regulates melanin production and controls melanocyte proliferation and apoptosis, providing a new theoretical basis for studying the mechanism of mammalian fur color formation.

黑素细胞在哺乳动物皮毛颜色的形成中起着重要作用,并受到几个基因的调节。尽管在黑色素瘤的研究中发挥了关键作用,但NRAS(神经母细胞瘤RAS病毒癌基因同源物)在哺乳动物表皮颜色形成中的机制仍是未知的。首先,采用qRT-PCR和Western blot检测不同颜色獭兔背部皮肤中NRAS mRNA和蛋白的表达水平。然后,用间接免疫荧光法鉴定NRAS在黑素细胞中的亚细胞定位。接下来,在黑素细胞中过表达和敲低NRAS的表达,并通过qRT-PCR和Western blot验证其有效性。随后用NaOH、CCK-8和Annexin V-FITC验证黑素细胞中黑色素含量、增殖和凋亡的变化。最后,我们分析了NRAS对其他影响黑色素生成的基因(MITF、TYR、DCT、PMEL和CREB)的调控。计算机研究表明,NRAS是一种稳定的亲水性蛋白,存在于黑色素细胞的细胞质和细胞核中。不同颜色獭兔皮肤中NRAS mRNA和蛋白的表达水平存在显著差异,以黑色皮肤表达量最高(P NRAS影响黑色素细胞的增殖、凋亡和黑色素的产生)(P NRAS与黑色素相关基因明显存在(P NRAS可作为调节黑色素产生、控制黑色素细胞增殖和凋亡的基因)。为研究哺乳动物皮毛颜色形成机制提供了新的理论依据。
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