鸡传染性支气管炎病毒HH06分离株的鉴定及其S1基因的分子特征

从黑龙江省哈尔滨市某鸡场疑似鸡传染性支气管炎的发病蛋鸡肾组织病料中分离到1株病毒,命名为HH06。该分离毒株经SPF鸡胚传代、血凝试验、负染电镜检查以及动物回归试验,证实为IBV。用RT-PCR扩增得到了HH06株S1基因片段,经测序,并与国内外IBV参考毒株的相应基因进行序列比较分析。结果表明,HH06株S1基因由1 620个核苷酸组成,推导的多肽由540个氨基酸残基组成,推导的氨基酸裂解位点序列为5个连续碱性氨基酸HRRRR。HH06株S1基因序列与除LX4外的其他参考株相应基因的核苷酸及氨基酸序列同源性均较低,亲缘关系均较远。初步证实HH06株为不同于IBV疫苗株的一个新毒株。     

广东鸽源禽Ⅰ型副黏病毒的分子生物学特征

为了解广东地区鸽源禽Ⅰ型副黏病毒的分子生物学特征,选择从该地区分离到的4株鸽源禽Ⅰ型副黏病毒(SD-55株、JM-11株、SZ-14株、ZQ-17株)进行了F基因的扩增及序列测定。结果表明,这4株病毒的F蛋白裂解位点均符合禽Ⅰ型副黏病毒强毒株裂解位点氨基酸序列的分子特征,与中国大陆鸽源APMV-1分离株Pigeon/YN-P1相比,相似率在95%以上。将其与GenBank中的18株鸽源禽Ⅰ型副黏病毒的相应序列进行比较分析,并绘制系统进化树,发现分离株SD-55株、JM-11株、SZ-14株位于以意大利分离株Pigeon/Italy/1166/00为代表的欧洲进化分支,而ZQ-17株位于以美国分离株Pigeon/U.S.(TX)/98为代表的美洲进化分支,经鉴定以上4株病毒均为基因Ⅵb1型禽Ⅰ型副黏病毒。     

猪传染性胃肠炎病毒SY株的分离鉴定及S基因的序列分析

取沈阳地区某猪场表现为腹泻症状的病死仔猪小肠病料,无菌处理后接种在PK-15细胞上分离病毒。通过无限稀释法进行病毒纯化、病毒中和试验和RT-PCR检测,证实其为传染性胃肠炎病毒(TGEV),并命名为TGEV SY株。参照Gen Bank中TGEV序列设计引物,对SY株S基因进行RT-PCR扩增,克隆并测序,获得S基因组完全序列。将S基因序列和所编码的氨基酸序列与Gen Bank中登录的10个TGEV和1个PRCV S基因序列进行同源性和亲缘关系的比较分析。结果表明,SY株S基因核苷酸序列和编码的氨基酸序列与其他参考毒株的同源性分别为96.4%~99.2%和96.0%~97.9%;而与PRCV毒株的同源性分别为96.2%和96.6%。由系统进化树可知,SY株与国内的华毒株、TS株以及国外的Miller M60株位于进化树的同一分支,亲缘关系较近。总体来看,各毒株之间的S基因差异性不大,表明TGEV虽然处于不断遗传演化中,但不同毒株的S基因变异不是特别明显。     

番鸭呼肠孤病毒YB株μB蛋白的原核表达与生物信息学分析

为研究番鸭呼肠孤病毒YB株(MDRV-YB株)μB蛋白基因与其他多株参考毒株之间的遗传进化关系以及对该序列的结构功能预测和进行蛋白的原核表达,本试验通过RT-PCR方法从MDRV中特异性扩增μB基因,并构建重组质粒pET-YB-μB,经测序获得其完整的CDS区;通过DNAstar等软件对MDRVYB株μB基因进行生物学信息分析,预测其编码蛋白的结构功能;重组质粒转化E.coli BL21(DE3)感受态细胞,经IPTG诱导表达μB融合蛋白。结果显示,μB蛋白基因CDS区全长2031 bp,共编码676个氨基酸;μB蛋白分子式为C3217H5105N879O1014S27,不存在信号肽,可能含有4个N-糖基化位点、85个磷酸化位点,二级结构中α螺旋结构、β螺旋结构、β转角结构和无规卷曲的氨基酸残基分别占27.6%,53.3%,10%和9.1%;在核苷酸同源性方面MDRV-YB株μB基因与番鸭呼肠孤病毒ZJ2000M株同源性高达99.8%,与其他多株鸡源呼肠孤病毒、鸭源呼肠孤病毒和鹅源呼肠孤病毒同源性都很低,仅为67.3%~69.6%;进化树分析结果显示,MDRV-YB株与MW9710株处于最近的分支;SDS-PAGE和Western-blot分析结果显示,经IPTG诱导,重组质粒pET-YB-μB成功表达了大小约为99.1 ku的μB融合蛋白,并且该蛋白反应原性良好。本试验结果为MDRV的遗传进化和μB蛋白功能的研究奠定了基础,为进一步研究MDRV的致病机制提供了数据支撑。     

鸡传染性支气管炎病毒SC株S1基因的序列分析

用RT PCR扩增了鸡传染性支气管炎病毒 (IBV)SC株S1基因 ,连接到 pMD 18 T载体上 ,克隆后进行了核酸序列分析 ,证实SC株S1基因与基因库中收录的国外毒株H12 0和M 4 1的同源性较低 ,分别为 81.1%和 80 .0 % ,而与国内JX990 1株的同源性达到 91.3% ,初步证实国内存在IBV新毒株。     

H7N2禽流感病毒分离株血凝素蛋白全基因的序列分析

为进一步分析H7N2禽流感病毒(AIV)分离株血凝素(HA)基因的特性,参照已发表序列设计了1对引物,采用RT-PCR获得了1条约1.7 kb的DNA片段,测序后进行了同源性比较、HA基因系统发育进化树分析和氨基酸编码分析。结果表明,所测的2个分离株的HA基因全长1 664 bp,编码除信号肽以外的HA蛋白的全部544个氨基酸,其中包括HA1的323个氨基酸,HA2的221个氨基酸。2个分离株HA基因核苷酸序列的同源性为99.4%;与GenBank中AIV标准株A/Afri.Star./Eng-Q/983/79(H7N1)的同源性最高,分别为99.4%和99.0%;与美国A/Chicken/NewYork/13142-5/94(H7N2)株同源性很低(仅65.0%),而与以色列、意大利H7N2AIV的同源性较高(为96%~97%);2个分离株在HA基因进化树中均处于H7亚型AIV的欧亚群系分支内。推导氨基酸的序列分析表明,其HA蛋白裂解位点的氨基酸序列为-GR-GLF-,仅包含1个碱性氨基酸(R-)残基,符合低致病力AIV的基因特征。     

一株新的鸽源基因Ⅵ型新城疫病毒的分离鉴定

从进口鸽中分离到1株鸽Ⅰ型副黏病毒,经测定,其鸡胚平均死亡时间(MDT)为59．2 h,鸡脑内接种致病指数(ICPI)为1．92,属于速发型新城疫病毒。通过RT-PCR,扩增出了该分离毒株F基因的重要功能区片段,经测序分析,其裂解位点的氨基酸序列为112RRQKR117F,符合新城疫强毒裂解位点特征。用blastn和blastx分别搜索GenBank的核酸和蛋白数据库,发现它与2株鸽Ⅰ型副黏病毒具有99％的同源性,系统发育进化树分析为基因Ⅵ型。表明,该病毒为1株新的强毒力鸽源基因Ⅵ型新城疫病毒。     

无血凝性兔出血症病毒VP60蛋白分子特性的分析及其在大肠杆菌内的高效表达

分离出1株在4、25、37℃条件下均不能凝集人"O"型红细胞的兔出血症病毒(RHDV),命名为SQ-1株;采用RT-PCR扩增获得病毒主要结构蛋白VP60蛋白基因,并对目的基因进行测序及分析;将目的基因定向克隆入原核表达载体pET-30a的多克隆位点,在大肠杆菌Rosetta(DE3)中诱导表达,纯化重组蛋白,用Western-blot检测其反应原性。测序与同源性分析结果显示,该毒株属于RHDV抗原遗传变异株(RHDVa),所扩增的VP60基因的ORF为1 740bp,编码579个氨基酸残基,与世界上其他RHDV毒株的核苷酸序列的同源性为89.9%～98.0%,与已报道的无血凝特性的病毒株whn-1株及低血凝性的Rainham株的核苷酸序列的同源性分别为95.2%及90.8%。重组质粒经IPTG诱导后目的基因获得表达,重组蛋白与预期大小一致,分子质量约为60ku,主要以包涵体形式存在。Western-blot检测证实,纯化蛋白能够与RHDV阳性血清发生良好的杂交反应,说明该重组蛋白具有良好的反应原性。     

猪细小病毒Z株NS部分基因的扩增及序列分析

从猪细小病毒Z株提取基因组DNA ,利用PCR扩增部分基因 ,并对该扩增片段进行测序与分析。结果表明 ,扩增的NS基因长 330bp ,编码 10 9个氨基酸。氨基酸序列中含有猪细小病毒的重要保守序列 ,并有 1个潜在的糖基化位点NFSN。Z株NS基因与其他猪细小病毒Kresse、NADL2 2、NADL2 1株的核苷酸同源性分别为 99%、98%、98% ,氨基酸同源性均为 99%。     

兔出血症病毒Yaan株衣壳蛋白VP60基因的克隆和序列分析

采用RT-PCR方法扩增兔出血症病毒(RHDV)Yaan株衣壳蛋白VP60基因,将扩增片段克隆到pMD 18-T载体上,经酶切及PCR鉴定后测序。结果显示,VP60基因全长1 740 bp,编码579个氨基酸。将Yaan株与其他RHDV分离株进行比较,结果显示,核苷酸同源性为90.5%~97.5%,氨基酸同源性为94.3%~99.5%,氨基酸变异多发生在衣壳蛋白C、E区,表明毒株具有高度保守性;同时对Yaan株与国外标准株AST/89的VP60蛋白氨基酸变异及其亲水性、柔性区、抗原区和表面结构进行了比较分析。     

The Development of EU-Russian Relations

The EU＇s strategy for Russia since the end of the cold war has to a large extent failed, and the direct consequences are the currently deadlocked EU-Russia relationship. There are three reasons for this： the EU and Russia＇s respective strength and status relative to each other have been changing since the beginning of this century; the EU＇s strategy towards Russia has been more fragmenting in recently years; and the American factor. The EU-Russia relations could continue to be in poor shape in the years to come, and it is very likely sometimes even further worsening. But the two sides would refrain from extreme measures and prevent their relations from sliding into real cold war.     

Young American China Watchers＇ Views on China

In this paper, the author has made a thorough study over the new generation of American China Watchers. Most of the pages are devoted to a detailed comparison between the old generation and the young generation of American China Watchers regarding to their background, education, and views over China. At the end of the paper, the author also gives some policy suggestions for both the Chinese and U.S. government based on the findings in this study.     

The Balance Between Coercion and Accommodation

1. Introduction
On the morning of April 1st, 2001, a U.S. EP-3 surveillance aircraft was conducting an intelligence flight along the Chinese territorial border in the airspace south-east of China＇s Hainan Island. Two Chinese fighter jets intercepted the spy plane and followed its flight, as is considered ordinary international practice. For reasons unknown, the EP-3 airplane made a sudden maneuver and crashed into one of the Chinese planes. The Chinese plane dropped into the sea, the pilot missing. The U.S. spy plane was also severely damaged. It intruded into Chinese territorial space and landed on Hainan Island without official permission from the Chinese side. After landing, the spy plane and 24 crew on board were detained by the Chinese authorities. This air collision episode was the first of its kind in the China-U.S. relations and an urgent diplomatic crisis ensued.     

U.S. and East Asian Integration

the U.S. is directly impacting on the future of the East Asian community. Therefore, finding ways to get along with the U.S. is crucial to the speed, direction, configuration and character of the East Asian community. In this paper, the author has analyzed the interests of the United States in East Asia and its stands towards the East Asian integration. The author concludes that it is to the interest of the United States to make more efforts to further join in the East Asian integration. And East Asia should accept and welcome the American participation.     

CAFIU Honorary Vice-President Wang Zhizhen Visited Turkey,Sudan and Attended the Third China-Africa People's Forum

正~~     

CAFIU Delegation Visited Sri Lanka and Attended the Third China-Africa People's Forum in Sudan

正At the invitation of Sri Lanka-China Friendship Association(SLCFA)and Sudan Network for National Non-Governmental Organizations,a 5-member delegation led by CAFIU Deputy Secretary-General Liu Kaiyang visited Sri Lanka and attended the Third China-Africa People’s Forum held in Sudan from May 7 to 14,2014.As a member of this delegation,I had the great honor     

South Asian NGO Delegation Visiting China

正At the invitation of the Chinese Association for International Understanding(CAFIU),a 10-member NGO delegation composed of representatives from media,think tanks and NGOs from Bangladesh,India and Sri Lanka visited China from April 9 to 18.On the second day of arrival,the delegation visited Xicheng District Administrative Service Center.The center provides such services as     

Building A Platform for Understanding and Establishing Channels for Cooperation——Notes on the 2014 “Understanding & Cooperation” Dialogue

正From June 16 to 18,the 2014"UnderstandingCooperation"Dialogue and Forum on Rethinking Asia,co-hosted by CAFIU and FES and jointly organized with China Foundation for Peace and Development,was held in Tutzing,a beautiful and spacious manor located at 30km southwest of Munich.Nearly 60 delegates from more than 10Asian and European countries participated in the event,including China,Germany,Korea,Japan,     

The Second Sino-US Maritime Security Workshop Held in CIIS

正On September 25-26,2014,CIIS and Hoover Institution,Stanford University jointly held the Second Sino-US Maritime Security Workshop in Beijing.CIIS President Qu Xing and Abraham Sofall,senior research fellow with the Hoover Institution,codelivered the opening remarks.Representatives from China's Ministry of National Defense,     

俄白哈关税同盟的发展及其影响（英文）

In recent years, the Eurasian Economic Community has developed rapidly. Because of the special relationship between the SCO and the Customs Because Customs Union, the latter will inevitably exert some influence on the development of the former, but they are not fundamentally contradictory. Since the of the establishment of the Customs Union, the trade between China and the Customs Union members grows steadily.