拟穴青蟹线粒体COI基因序列克隆及SNPs位点分析

根据拟穴青蟹(Scylla paramamosain)线粒体全序列设计引物,采用PCR产物双向测序法,克隆了线粒体细胞色素C氧化酶亚基Ⅰ(COI)基因部分序列(761 bp),并筛选、分析了拟穴青蟹的SNPs位点.共分析了采自福建省宁德市的16只拟穴青蟹,另外,从GenBank数据库中下载了31条前人提交的拟穴青蟹COI基因序列(长度在425-522 bp之间).利用MEGA 4.0软件对所有序列进行比对分析,结果表明碱基T、C、A和G的平均含量分别为37.6％、17.8％、28.9％和15.7％,G+C的含量平均为33.5％.分析结果表明,在所克隆的761bp的COI基因序列中共发现29个SNPs位点,其出现频率为3.8％.在这29个SNPs位点中,13个为C/T转换(占44.8％),12个为A/G转换(占41.4％),2个为A/T颠换(占6.90％),1个为G/C颠换(占3.45％),另有一个位点(201 bp处)发生了A/C颠换和C/T转换两种类型的碱基替换.氨基酸分析表明,在29个SNPs位点中,有9个位点属于非同义突变,引起了氨基酸的变化;其他20个位点属于同义突变,未引起氨基酸的变化.这将为拟穴青蟹遗传背景和群体遗传多样性研究提供新的分子标记.     

中国水域瓶鼻海豚的mtDNA控制区序列变异性分析

测定了30头中国水域瓶鼻海豚（Tursiops sp.)mtDNA控制区5′端424bp的序列,结合已发表的中国水域其它瓶鼻海豚的mtDNA控制区序列,共发现54个变异位点,定义了37种单元型。中国水域瓶鼻海豚的两个形态型之间没有共享单元型,且具有8个鉴别位点。基于最大似然法和邻接法的系统发生分析均把单元型聚类为分别代表两个形态型的支系。形态型之间的核苷酸歧异度为5.58%,超过了其它海豚类种间的序列歧异水平,支持把这两个形态型划分为两个独立的种,即T.truncatus和T.aduncus的观点。虽然两种瓶鼻海豚的分布区在台湾海峡一带出现重叠,但相互之间缺乏基因流动,提示两者可能已出现了显著的生殖隔离。     

基于线粒体细胞色素c氧化酶亚基I基因序列的帘蛤科贝类分子系统发育研究

对21种帘蛤科贝类线粒体细胞色素c氧化酶亚基Ⅰ(cytochrome c oxidase subunit I,COI)基因核苷酸序列进行了分析,以探讨这一序列在种质鉴定、分子系统发生研究中的应用价值。测序结果表明,所有物种扩增片段长度均为707 bp(含引物),序列A+T含量(62.4%—67.8%)明显高于G+C含量。物种间共有变异位点379个,其中简约信息位点334个;此区段共编码235个氨基酸,种间共有氨基酸变异位点100个。以COI基因片段序列为标记,用中国蛤蜊(Mactra chinensis)作外群,构建了35种帘蛤科贝类(其中14种贝类COI序列从GenBank下载)的系统发生树,结合拓扑结构分析和序列比对分析,结果表明:支持将短文蛤(Meretrix petechinalis)和丽文蛤(M.lusoria)订为文蛤(M.meretrix)的同物异名的观点,建议将丽文蛤和短文蛤订为文蛤的地理亚种;支持将薄片镜蛤(Dosinia corrugata)和D.angulosa订为2个独立种的观点;认为将波纹巴非蛤(Paphia undulata)和织锦巴非蛤(P.textile)订为2个独立种是合适的。COI基因序列含有丰富的遗传信息,适合作为帘蛤科贝类种群遗传结构和系统发生研究的分子标记。     

雷州地区马尾藻系统进化分析

通过PCR扩增测序及种间比对,对获得的6种马尾藻18SrRNA、COI和ITS基因部分同源序列进行分析,结果显示:(1)18S rRNA基因同源序列长度为1 673 bp,其中保守位点1 668个,可变位点3个,简约信息位点1个,单变异多态位点2个;T、C、A、G的平均含量分别为26.4％、21.4％、24.4％、27.8％.(2)COI同源序列长度为643 bp,其中保守位点567个,可变位点76个,简约信息位点28个,单变异多态位点47个;T、C、A、G的平均含量分别为40.0％、17.6％、19.1％、23.3％.(3)ITS同源序列长度为1 539 bp,其中保守位点1 272个,可变位点193个,简约信息位点55个,单变异多态位点138个;T、C、A、G的平均含量分别为23.5％、26.5％、17.1％、32.9％.(4)基于Kimnra双参数模型计算遗传距离,选用褐藻纲部分物种为外源,构建18S rRNA,COI和ITS基因序列系统发育树.用COI和ITS基因构建的发育树与利用形态学进行的分类较为一致,真马尾藻亚属(Sargassum)的张氏马尾藻(Sargassum zhangii)、全缘马尾藻(Sargassum integerrimum)、匍枝马尾藻(Sargassum polycystum)、亨氏马尾藻(Sargassum henslowianum)、灰叶马尾藻(Sargassum cinereum)亲缘关系较近,先聚为一支,再与反曲叶亚属的半叶马尾藻(Sargassum hemiphyllum)聚为一支,然后与其他亚属的聚为一大支;而用18S rRNA基因构建的发育树中,亨氏马尾藻先与半叶马尾藻聚为一支,后与真马尾藻亚属的聚为一支,最后与其他亚属的聚为一大支.从序列的保守度分析,18SrRNA基因具有高度保守性,可用于属以上单元鉴别分类;COI和ITS基因多态性较高,可用于种间分类.     

林麝MHC-DRA基因分析及与反刍亚目DRA基因比较

MHC-DRA基因仅在少部分物种如马、斑马、驴等具有多态性,DRA基因在其他哺乳动物中多为低多态性。由于MHC-DRA的低多态性,较少受到研究者重视,我们选取近缘物种牛的DRA基因引物,对四川养麝研究所的217只林麝进行DRA基因exon 2的序列扩增,得到2个林麝DRA基因,片段长度为260 bp,这2个基因仅有一个核苷酸同义突变,没有氨基酸突变。我们将所获得的林麝DRA基因与反刍亚目其他物种DRA基因进行氨基酸比对发现,我们扩增的序列为林麝DRA基因exon 2,编码MHCⅡ类分子的α1区域,而这段区域主要参与形成MHCⅡ类分子的多肽结合槽,根据Brown确定人类的抗原肽结合区位点,在参与比对的反刍亚目物种DRA基因中,抗原位点11、22、72、76存在氨基酸变异,而在这些位点上林麝的DRA基因没有变异,和大部分反刍亚目相同。同时我们发现大部分反刍亚目物种之间氨基酸突变的位点大都在非抗原结合位点。     

两头人工饲养南瓶鼻海豚的病理解剖及死因分析

<正>南瓶鼻海豚(Tursiops aduncus)是我国二级重点保护的水生哺乳动物,属于鲸目(Cetacea),海豚科(Delphinidae),瓶鼻海豚属(Tursiops)(Mller and Beheregaray,2001;Wells and Scott,2002)。南瓶鼻海豚是人工主要饲养的鲸类品种之一(刘仁俊等,2002;Zhang et al.,2012)。2011年4月30日,厦门市小嶝岛休闲渔村从福建东山引进两头南瓶鼻海豚进行人工饲养。饲养池位于北     

基于13 个内含子的序列探讨鲸目的系统发育关系

本文基于实验室筛选得到的13 对内含子标记,在鲸偶蹄目的15 个物种中进行有效扩增,并重建了这15
个物种的系统发育关系。结果表明,抹香鲸总科(Physeteroidea) 位于齿鲸亚目(Odontoceti)的基部,从而支
持了传统的齿鲸亚目的单系性。在海豚总科(Delphinoidea)内部,贝斯分析结果支持了鼠海豚科(Phocoenidae)
和一角鲸科(Monodontidae)的姐妹群关系,而后再与海豚科(Delphinidae)相聚。系统发育分析同时还
强烈支持了海豚科的四个属(Sousa,Tursiops,Stenella,Delphinus)组成一个单系的“复合体”。另外,我们的分
析结果并不支持瓶鼻海豚属(Tursiops)和原海豚属(Stenella)的单系性。基于松散分子钟的分歧时间估算与以
往文献中的结果没有明显差异。这些研究结果提示,核基因内含子序列有希望解决一些长期存在的鲸类系统发
育问题。     

东亚三角涡虫Rab蛋白cDNA序列的克隆与生物信息学分析

通过构建东亚三角涡虫(Dujesia japonica)cDNA文库,随机挑选重组阳性克隆进行测序,对部分序列进行引物步移法测序,获得1个三角涡虫新基因——Rab蛋白基因(DjR),涡虫Rab蛋白cDNA全长2 141 bp,开放性阅读框(ORF)621bp,编码206个氨基酸,相对分子量为23.1 kD,等电点6.59,属亲水性蛋白,主要定位于细胞质中,在氨基酸第20和21位之间有信号肽剪切位点。有8个磷酸化位点。含有小G蛋白家族5个保守的鸟苷酸结合区域。同源性比较分析结果表明,其碱基序列与已经报道的其他23个物种的相似性为53%-90%,且符合种属之间的进化关系。     

百合psbA-trnH基因序列变异及遗传多样性分析

目的:通过对叶绿体基因psbA-trnH序列变化差异来研究百合遗传多样性。方法:利用PCR技术扩增百合叶绿体ps-bA-trnH基因片段并测序,再用生物软件Clustalx2、Mega5.0、DnaSP对测序结果进行分析。结果:扩增的psbA-trnH基因片段长度约为469～583bp,GC含量为34.47～35.20%,其核苷酸变异位点27个,信息位点12个,变异位点主要集中在88～107 bp区域,根据这些变异位点供试材料可以分为3类,核苷酸的多样性为0.01150。结论:叶绿体基因trnH-psbA序列可以区分百合种间的差异,可以作为一种很好方法来研究遗传多样性。     

金花茶查尔酮合成酶基因全长克隆与序列分析

利用RT-PCR和RACE方法,从我国珍稀植物金花茶(Camellia nitidissima)花瓣中获得了查尔酮合成酶(chalcone synthase,CHS)基因的cDNA全长,命名为Cn-CHS,GenBank登录号HQ269804.碱基序列分析表明,Cn-CHS全长1 454bp,包含77 bp的5'非翻译区、207 bp的3'非翻译区和一个长为1 170 bp编码389个氨基酸的开放阅读框.氨基酸序列分析显示该基因编码的蛋白具有CHS家族保守存在的所有功能活性位点和特征性多肽序列.氨基酸序列比对分析表明,CnCHS与蔷薇科、杜鹃花科、茄科等植物的CHS相似性都在92%以上;与山茶科山茶属物种山茶(C.japonica)CHS完全一致;与茶(C.sinensis)CHS相似性达99%,有5个氨基酸位点存在差异,其中包括一个功能性位点.     

A Unique Protease Cleavage Site Predicted in the Spike Protein of the Novel Pneumonia Coronavirus (2019-nCoV) Potentially Related to Viral Transmissibility

正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases     

Curcuminoids as inhibitors of thioredoxin reductase: A receptor based pharmacophore study with distance mapping of the active site

Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable potent antiproliferative synthetic drugs.     

Comparative morphology of the carpel in the Liliaceae: Hewardieae, Petrosavieae, and Tricyrteae

The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle.     

Enterovirus 71 3C proteolytically processes the histone H3 N-terminal tail during infection

Highlights
1. The N-terminal tail of histone H3 is specifically cleaved during EV71 infection.
2. Viral protease 3C is identified as a protease responsible for proteolytically processing the N-terminal H3 tail.
3. Our finding reveals a new epigenetic regulatory mechanism for Enterovirus 71 in virus-host interactions.     

Detection of EBV and HHV6 in the Brain Tissue of Patients with Rasmussen’s Encephalitis

Rasmussen’s encephalitis (RE) is a rare pediatric neurological disorder, and the exact etiology is not clear. Viral infection may be involved in the pathogenesis of RE, but conflicting results have reported. In this study, we evaluated the expression of both Epstein-Barr virus (EBV) and human herpes virus (HHV) 6 antigens in brain sections from 30 patients with RE and 16 control individuals by immunohistochemistry. In the RE group, EBV and HHV6 antigens were detected in 56.7% (17/30) and 50% (15/30) of individuals, respectively. In contrast, no detectable EBV and HHV6 antigen expression was found in brain tissues of the control group. The co-expression of EBV and HHV6 was detected in 20.0% (6/30) of individuals. In particular, a 4-year-old boy had a typical clinical course, including a medical history of viral encephalitis, intractable epilepsy, and hemispheric atrophy. The co-expression of EBV and HHV6 was detected in neurons and astrocytes in the brain tissue, accompanied by a high frequency of CD8⁺ T cells. Our results suggest that EBV and HHV6 infection and the activation of CD8⁺ T cells are involved in the pathogenesis of RE.     

Perinatal Vertical Transmission of Chikungunya Virus in Ruili,a Town on the Border between China and Myanmar

正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016).     

Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Method for Rapid Detection of SARS-CoV-2

In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans.