番木瓜环斑病毒畸叶株系的CP基因克隆和序列分析

采用RT-PCR方法合成了本研究室保存的番木瓜畸叶病毒(PMaLV)的外壳蛋白(CP)基因,将其CP基因克隆进Promega公司的pGEM-T and pGEM-T Easy Vector System(简称T-载体),并进行了序列分析.结果表明,PMaLVCP基因核苷酸序列全长为861nt,推导其编码287个氨基酸.与番木瓜环斑病毒(PRSV)美国夏威夷HA株系和澳大利亚W株系的CP基因相比,在第66nt处开始连续缺失3个核苷酸.与PRSV的华南Ys、Sm和G株系以及夏威夷的HA和澳大利亚的W株系相比,其CP基因序列同源率分别为96%、98%、95%、89%和89%.其推导的氨基酸序列同源率分别为98%、97%、97%、96%和95%.此结果表明,PMaLV属于PRSV的一个株系,不是一种新病毒.因此,我们称其为番木瓜环斑病毒畸叶株系(ML株系).     

大蒜A病毒CP基因的原核表达及抗血清制备

根据GenBank报道的大蒜A病毒(GarV-A)序列设计引物、扩增其外壳蛋白基因并进行序列分析.结果表明,GarV-A的CP基因与目前已报道的两种GarV-A不同分离物CP基因的核苷酸序列同源性为98％-99％;氨基酸序列同源性均为98％.将GarV-A CP基因插入表达载体pSBET,在大肠杆菌BL21(DE3)Plys E菌株中诱导表达.CP经12％SDS-PAGE和5％-20％SDS-PAGE两次纯化,免疫小鼠获得抗CP血清,Western blotting分析表明确定制备的抗体对CP具有高度特异性,ELISA检测表明制备的抗体能够与天然病毒离子结合,因此可以作为该病毒的检测.     

桃拉综合征病毒中国株ZHZC3全基因测序及分子结构预测

设计8对引物分片段扩增桃拉综合征病毒中国分离株ZHZC3全基因组,病毒两末端序列采用末端快速扩增方法(RACE)获取.扩增产物克隆到pMD18-T载体并测序,用DNAstar软件拼接全序列及同源性比较.结果显示ZHZC3全序列除去3' poly (A)尾,由10202个碱基组成,有两个开放阅读框,分别编码2107和1011个氨基酸的聚蛋白.与美国参考株HI94相比,在编码区没有核苷酸的缺失和插入,但在5' UTR缺失3个A,两者整体核酸同源性达97.9%.ORF1 中ZHZC3与HI94及巴西株(BLZ01)的核酸同源性分别为97.6%、97.7%,在ORF2中ZHZC3与HI94、BLZ01的核酸同源性则分别为98.3、97%.与国外株ORF2的部分序列比较发现ZHZC3和中国台湾株均与美国株HI94同源性最高.克隆分析6株TSV中国大陆株主要结构蛋白CP2基因,发现其编码的氨基酸存在三个高变区,中国大陆株更有其独特的氨基酸变异模式,312 (S), 449 (A), 451 (Q) 和468 (H).表明该病毒的整体变异性不高,但中国的流行株已形成其自己的遗传演变特征.在此基础上,利用生物学软件对CP2蛋白功能域和三维结构进行了预测,为进一步分析CP2蛋白结构与功能关系奠定了基础.ZHZC3株是第一个测定全序列的TSV中国株.     

中国大豆花叶病毒SC7外壳蛋白基因的序列测定及其与美国株系的比较北大核心CSCD

为了探索大豆花叶病毒（Soybean mosaic virus,SMV）SC7外壳蛋白（Coat protein,CP）基因序列与病毒致病性作用及其与美国株系的对应关系,本研究通过反转录-聚合酶链式反应（RT-PCR）,克隆了我国SMV SC7株系的CP基因,并测定了其全序列。结果表明：CP基因长度为795bp,编码的CP蛋白为265个氨基酸。在致病性反应上,SC7的毒力比美国的毒株更强些。在分子水平上,CP基因核苷酸序列差异为4%~5%,编码的氨基酸序列差异为1%~2%。美国株系N端都存在与蚜传有关的保守基序DAG（Asp-Ala-Gl）,但在SC7为DAD（Asp-AlaAsp）。将序列信息与SMV株系在鉴别寄主上的症状反应进行综合分析发现,CP基因的同源性与SMV在鉴别寄主上的致病性反应没有明显关系。     

中国大豆花叶病毒SC7外壳蛋白基因的序列测定及其与美国株系的比较

为了探索大豆花叶病毒(Soybean mosaic virus,SMV)SC7外壳蛋白(Coat protein,CP)基因序列与病毒致病性作用及其与美国株系的对应关系,本研究通过反转录-聚合酶链式反应(RT-PCR),克隆了我国SMV SC7株系的CP基因,并测定了其全序列。结果表明:CP基因长度为795bp,编码的CP蛋白为265个氨基酸。在致病性反应上,SC7的毒力比美国的毒株更强些。在分子水平上,CP基因核苷酸序列差异为4%~5%,编码的氨基酸序列差异为1%~2%。美国株系N端都存在与蚜传有关的保守基序DAG(Asp-Ala-Gl),但在SC7为DAD(Asp-AlaAsp)。将序列信息与SMV株系在鉴别寄主上的症状反应进行综合分析发现,CP基因的同源性与SMV在鉴别寄主上的致病性反应没有明显关系。     

由高粱花叶病毒和甘蔗花叶病毒引发的浙江甘蔗花叶病害

从浙江省5个地区采集表现花叶症状的甘蔗病叶,用马铃薯Y病毒科简并引物做PCR扩增及测序鉴定.序列分析表明,5个甘蔗样品均含有高粱花叶病毒(SrMV),其中3个样品中还存在甘蔗花叶病毒(SCMV)的复合侵染.序列比较和系统进化树分析表明,浙江甘蔗样品中的SrMV序列彼此很相似,核苷酸同源性大于93%,与已报道的4个美国分离物在CP区域同源性很高,但是3′非编码区的同源性却仅为70%左右.SCMV欧洲和中国玉米分离物及美国、南非和澳大利亚甘蔗分离物分别形成两个远缘群体,浙江甘蔗分离物群体位于两者之间;群体间CP氨基酸序列同源性均大于80%.甘蔗和玉米上的SCMV差异明显,多为无义突变.     

芜菁花叶病毒的3’末端序列克隆与CP基因序列分析

目的:克隆芜菁花叶病毒(Turnip mosaic virus,TuMV)的3'末端序列,并进行CP基因序列分析.方法:以TuMV杭州榨菜分离物(TuMv-HZZC)接种病叶为材料,利用病毒粒子吸附法制备病毒RNA模板,经RT-PCR扩增获得了TuMV-HZZC 3'末端序列,将其克隆到PMD 18-T质粒上进行序列分析.结果:TuMV-HZZC分离物3'末端序列包括部分的Nib基因、完整的TuMVCP基因和3'-UTR,CP基因为864bp,分别编码288个氨基酸,3'-UTR序列(不包括PolyA尾巴)为213bp.经过与其他TuMV分离物的CP基因核苷酸和氨基酸比较,同源性分别达到88.0%～97.6%和91.0%-96.5%.结论:TuMV的系统进化具有典型的地域和寄主关联性.     

小西葫芦黄化花叶病毒分离物的3′末端序列多态性研究

研究了来自中国大陆9个小西葫芦黄化花叶病毒（ZYMV）分离物的基因组3′末端核苷酸序列及所推导的外壳蛋白（CP）氨基酸序列以及3′末端非编码区（UTR）序列,并与其它地区所报道的16个ZMYV分离物进行了同源性比较。ZYMV CP基因核苷酸序列具有一定的寄主相关性和地域相关性,但总体上其关联程度不明显;同时,CP氨基酸序列的寄主适应性程度明显高于地域相关性。25个ZYMV分离物的CP氨基酸序列根据其变异程度分为2个区： N端约41个氨基酸为高度变异区,CP核心区和C端氨基酸序列为保守区。研究结果初步揭示了ZYMV作为单链RNA病毒通过与寄主相互作用而表现寄主适应性变异的趋势。     

大豆花叶病毒CP基因原核表达与抗血清制备

根据已报道的大豆花叶病毒(SMV)序列设计引物,扩增SMV的外壳蛋白(CP)基因,序列同源性分析结果表明,与目前已报道的SMV不同分离物CP基因核苷酸序列同源性为85.0%～96.8%,相应推测的氨基酸序列同源性为87.9%～98.9%.将CP基因插入原核表达载体pSBET后在大肠杆菌BL21(DE3)Plys S中诱导表达.通过12%SDS-PAGE和5%～20%梯度SDS-PAGE两次制备电泳纯化诱导产物,免疫家兔获得抗CP血清,Western blot分析对CP具有高度特异性.硫酸铵沉淀法与Protein A-Red Sepharose 亲和层析相结合提取IgG,获得效价达1∶ 3 800的一抗,可用于田间SMV病样检测.     

番木瓜环斑病毒畸叶株系的CP基因克隆和序列分析

采用RT－PCR方法合成了本研究室保存的番木瓜畸叶病毒（PMaLV）的外壳蛋白（CP）基因,将其CP基因克隆进Promega公司的pGEM-T and pGEM-T Easy Vector System（简称T－载体）,并进行了序列分析。结果表明,PMaLV CP基因核苷酸序列全长为861nt,推导其编码287个氨基酸。与番木瓜环斑病毒（PRSV）美国夏威HA株系和澳大利亚W株系的CP基因相比,在第66nt处开始连续缺失3个核苷酸。与PRSV的华南Ys、Sm和G株系以及夏威夷的HA和澳大利亚的W株系相比,其CP基因序列同源率分别为96％、98％、95％、89％和89％。其的氨基酸序列同源率分别为98％、97％、97％、96％和95％。此结果表明,PMaLV属于PRSV的一个株系,不是一种新病毒。因此,我们称其为番木瓜环斑病毒畸叶株系（ML株系）。     

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.