北极狐白介素-18基因的克隆与序列分析

根据GenBank中登录的犬白介素-18(IL-18)基因序列,设计了1对特异性引物.将北极狐外周血淋巴细胞经植物血凝素(PHA)诱导,用TRIzol裂解,提取总RNA,通过反转录聚合酶链式反应(RT-PCR)扩增狐狸IL-18基因,连接pMD18-T载体,转化DH5α感受态细胞,获得阳性重组质粒.核苷酸测序结果显示,获得的基因序列全长590 bp,含582 bp的开放阅读框,编码193个氨基酸,与已知犬IL-18序列的同源性高达98.7%,氨基酸同源性为96.4%.     

猪三种细胞因子基因的克隆与序列测定

利用RTPCR技术,从植物凝集素(PHA)、脂多糖(LPS)诱导的猪外周血淋巴细胞(PBMC)中成功地克隆了IL2、IL4、IFNα1基因,诱导后第6～24h细胞因子转录基因量最丰富;诱导剂LPS和PHA联合使用的效果优于LPS或PHA单独使用的诱导效果。序列测定与分析表明,克隆的IL2、IL4和IFNα1基因与相应的参考序列的同源性分别是98.5%、99.0%和97.6%。     

鹅IL-18基因的克隆和原核表达及其多克隆抗体的制备

参考GenBank中鸡、火鸡和鸭IL-18基因序列,设计了1对特异引物,以提取的鹅脾细胞总RNA为模板,应用RT-PCR扩增、克隆鹅IL-18基因中间的333bp序列。序列分析结果表明,其与鸭IL-18基因对应序列相似性最高,达99%。参考鸭IL-18全基因序列设计上、下游引物各3条,以梯度PCR探索扩增、克隆鹅IL-18全基因。以上述策略成功扩增、克隆了3个广东地方杂交品种鹅的IL-18基因。序列分析表明,这3个杂交品种鹅的IL-18基因序列一致,长642bp,含一个603bp的ORF,编码200个氨基酸,分子质量为23.2ku。ORFN端的30个氨基酸为前导序列,其余170个氨基酸为功能蛋白序列。在GenBank中注册了第一条鹅的IL-18基因序列,登录号为EF159728。随后,构建了鹅IL-18基因ORF的重组原核表达质粒pET-GIL-18,经诱导表达、纯化获得重组鹅IL-18融合蛋白,并以其制备了兔抗重组鹅IL-18多克隆抗体。     

鸡白细胞介素5基因的克隆与原核表达

根据GenBank中登录的鸡白细胞介素5(IL-5)基因序列,设计并合成1对特异性引物,首次从ConA激活的鸡脾淋巴细胞中扩增出了鸡IL-5基因cDNA。序列分析表明,该基因全长381bp,编码126个氨基酸,蛋白质分子质量约14.63ku,包括22个氨基酸长度的信号肽。与其他哺乳动物IL-5的氨基酸同源性低于25%,显示较强的种属特异性。但该蛋白的三维结构与人IL-5蛋白结构相似,主要由4个α螺旋构成。进而构建了此基因的原核表达载体pET-28a-IL-5,并在大肠杆菌Rosetta(DE3)中通过IPTG的诱导成功实现了表达,为进一步研究禽IL-5的结构和生物学功能奠定了良好的基础。     

杜氏利什曼原虫NH36基因的克隆及其在大肠杆菌中的表达

应用RT-PCR技术扩增并克隆杜氏利什曼原虫(Leishmania donovani)NH36编码基因,经鉴定及序列分析,构建其原核重组表达载体,经IPTG诱导表达,SDS-PAGE和Western-blotting鉴定其表达产物。结果显示,获得的NH36开放阅读框为945 bp,编码314个氨基酸残基,与GenBank上发表的国际标准株NH36编码基因序列以及氨基酸序列同源性分别为88.57%(837/945)和89.17%(280/314)。表达蛋白为36 ku,经1 mol/L IPTG诱导6 h后的表达量最高;薄层扫描显示表达的蛋白占菌体蛋白总量的57%;该蛋白可被抗杜氏利什曼原虫的多克隆抗体血清识别。     

鸭白细胞介素-2基因在毕赤酵母中的表达

根据GenBank中登录的鸭IL-2基因序列设计并合成了1对特异性引物,以经ConA诱导的广州鸭外周血淋巴细胞提取的总RNA为模板,用RT-PCR方法扩增出长度为360 bp的目的基因片段,并将该基因克隆到pMD18-T载体上.酶切鉴定、PCR鉴定及序列测定结果表明,获得了鸭IL-2成熟蛋白基因的完整克隆.测序结果表明,该成熟蛋白基因由360个核苷酸组成,共编码119个氨基酸.克隆的鸭IL-2基因与GenBank上序列号为AYl73028及AF294322的鸭IL-2基因的同源性高达99.4%.将目的基因克隆至真核表达载体pPICZáC上,构建了重组质粒pPICZáC-DuIL-2.酵母转化子经甲醇诱导发酵分泌表达了鸭IL-2基因.SDs-PAGE分析证实,鸭IL-2基因在毕赤酵母(Pichia pastoris)中表达成功,表达的重组蛋白的分子质量约为14.3 ku.     

痢疾杆菌福氏2a及其环丙沙星耐药株gyrA和parC基因耐药性决定区序列的测定分析

测定了痢疾杆菌福氏 2a 30 1株与喹喏酮类药物耐药性相关的 gyrA基因和 parC基因的序列 ,并对其环丙沙星耐药诱变株的 gyrA和 parC基因喹喏酮类药物耐药性决定区 (QRDR)序列进行了测定分析。结果表明 ,痢疾杆菌福氏 2a 30 1株 gyrA和 parC基因分别为 2 6 2 5bp和2 2 5 6bp ,环丙沙星诱导的耐药菌 gyrA基因QRDR(2 4 5bp)发生氨基酸残基 6 9 Ala→Val和 87 Tyr→Asp改变 ,parC基因QRDR(2 37b)发生氨基酸残基 79 Ala→Asp、84 Ala→Glu和 85 Pro→Ala改变。这一研究结果对认识痢疾杆菌喹喏酮类药物耐药性的分子机理具有重要意义     

双峰驼朊蛋白基因的克隆和序列分析

分别从4峰双峰驼全血中提取基因组总DNA,用所设计的引物扩增了细胞型朊蛋白(PrP)基因,并克隆到pMD 18-T载体。序列分析表明,所克隆的4个双峰驼PrP基因片段大小分别为768、768、792和795 bp,包含了朊蛋白基因的完整编码区序列,为包含在单一外显子内的完整开放阅读框,均与国外报道的同属单峰驼PrP序列基本相同。4个双峰驼PrP基因含5个或6个短而富含G-C的元件,可编码5个或6个八肽(九肽)重复序列Pro-His-Gly-Gly-Gly-Trp-Gly-Gln或Pro-Gln/His-Gly-Gly-Gly-Gly-Trp-Gly-Gln,其氨基酸序列含有24个氨基酸的N-端信号肽和22个氨基酸(除LT200302为23个氨基酸外)的C-端信号肽。4个双峰驼PrP基因之间相比较,其核苷酸和推导氨基酸序列的同源性为91.0%~100.0%和94.2%~100.0%。共发生133个碱基替换,其中107个为同义码替换,26个为异义突变。     

新城疫病毒广东分离株HN基因的克隆与序列分析

用RT PCR方法从6个新城疫病毒(NDV)广东分离株中扩增HN基因cDNA片段,并将其克隆至pGEM T Easy载体进行核苷酸序列测定。结果表明,6个NDV分离株 HN基因片段长度均为1 704 bp,编码568个氨基酸;彼此间核苷酸和氨基酸同源性分别为 96.0%~99.8%和98.6%~100%,与其他基因Ⅶ型毒株的氨基酸序列同源性为 96.8%~98.4%;但与其他基因型毒株如D26、Ulster/67、B1、LaSota以及GB Texas的氨基酸同源性较低,为88.2%~91.0%。     

广西10个地方优质品种鸡γ-干扰素基因的克隆与序列分析

根据基因库中鸡γ-干扰素的基因序列设计了1对特异性引物,应用反转录-聚合酶链式反应(RT-PCR)技术,从ConA诱导培养的广西10个地方优质品种鸡外周血淋巴细胞RNA中扩增了γ-干扰素基因,结果。均得到了大小为520 bp的特异性片段。将扩增产物纯化并克隆到 pMD18-T载体上,获得重组质粒,经PCR和EcoR Ⅰ Sal Ⅰ双酶切鉴定后测序。序列分析结果表明,广西10个地方优质品种鸡γ-干扰素基因均编码145个氨基酸的成熟蛋白,分子质量约为16．8 ku,与哺乳动物和其他品种鸡的核苷酸序列同源性分别为38．8％～39．0％和99．2％～100％,氨基酸序列同源性分剐为23．6％～24．8％和97．6％～99．4％。     

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.     

中国需要构建怎样的周边(英文)

正The new central leadership has highlighted the strategic importance of better relations with neighboring countries and the construction of a solid foundation for China’s neighborhood strategy so as to create favorable conditions for Chinese development.1China is strengthening top-level strategy in foreign relations,focusing on global-level relations and staying committed to relations     

Learning from Others and Starting Built-to-last Companies

正Ruicheng Group was founded in Qiqihar,a city located in one of the three major ‘Chernozem belts’ in the world,the fertile Songnen Plain,and one known as the ‘home of red-crowned cranes’ Over the years,our business developed from being only one supermarket in the beginning to a corporate group operating in multiple sectors,from supermarket chain and real estate to property management and preschool education.     

中巴关系的发展及其认识误区（英文）

China and Brazil established a strategic partnership in 1993. The bilateral ties have seen rapid development in almost all areas. However, there are eight myths surrounding the China-Brazil relations. These myths are the result of a lack of mutual understanding as well as the misconceptions result misconceptions of the western observers and media. These misunderstandings have of have jeopardized images of the China-Brazil relations and also hindered further development of their bilateral relationship.     

中国的“主场外交”:机遇、挑战与任务（英文）

The successful hosting of the CICA summit has drawn great attention The attention to China's "host diplomacy". Analyzing new features of China's "host to "host diplomacy" during this new period, this paper will look at the opportunities and challenges facing China's "host diplomacy" and discuss the problems that China should address in order to do a better job in this regard and bring its "host diplomatic advantages" into full play.     

亚洲整体性崛起及其效应(英文)

Since the beginning of the 21st century, the overall rise of the Asian geographic plate has been particularly prominently. The overall rise of Asia reflects the improvement of a large number of developing countries in Asia. The long-occupied aggressive status of the United States and Europe is weakening, and the overall rise of Asia appears more highlighted, and its impact upon the world seems more intense.     

CAFIU Convenes the 11~(th) Council Meeting in Beijing

正On the morning of April 2,2014,CAFIU held the 11th Executive Council Meeting in Beijing.Present at the Meeting were Member of the Politburo of the 17th CPC Central Committee and Vice-Chairman of the 11th CPPCC National Committee Mr.Wang Gang,ViceChairman of the 11th NPC Standing Committee and President of CAFIU Mr.Zhou Tienong,ViceChairpersons of the NPC Standing Committee Mme     

Speech by Honorary President Zhou Tienong at the 11~(th) Executive Council Meeting of CAFIU (Excerpt)

正I am very pleased to attend the 11th Executive Council Meeting of CAFIU,and would like to express my hearty congratulations to President Yan Junqi,the new leadership and the newly-elected Executive Council.I also want to express my sincere thanks for choosing me to be the Honorary President of CAFIU!