猪带绦虫TSO18基因真核表达载体的构建及其表达

采用PCR从重组克隆载体pGEMTSO18中扩增出猪带绦虫六钩蚴TSO18基因,与毕赤酵母分泌性表达载体pPIC9k相连接,构建重组表达载体pPIC9kTSO18,转化大肠埃希氏菌JM109,经测序证实,基因序列完全正确。制备重组质粒pPIC9kTSO18,用SalⅠ线性化,并电转化毕赤酵母(Pichiapastoris)GS115,使重组表达载体与酵母染色体发生同源整合。采用G418抗性梯度法筛选得到高拷贝重组菌株,以甲醇进行诱导表达,SDSPAGE和Westernblotting分析结果表明,诱导表达的培养上清中表达出具有反应活性的16ku重组蛋白,目的蛋白约占培养上清液中蛋白总量的80%以上,诱导72h目的蛋白表达量为0.5mg/mL。     

县域城镇化发展模式探究——以河北省青龙县为例

青龙县在城镇化发展过程中既有优势，也存在不足。基于对青龙县政治、经济、技术、社会等因素分析，需要从历史和现实的角度出发。根据资源禀赋状况准确定位。要依托资源优势，强化产业支撑；建设中心城镇，完善城市功能；加快制度创新，改善城镇环境；加强城镇管理，提升服务质量，从而因地制宜地走出健康县域城镇化之路。     

Better Environment,Better Games

No one would believe that Donghaochong Stream,now a leisure site for Guangzhou's residents,used to be a smelly,stagnant waterway of filth; it was a stain on the city's image in the run-up to the Guangzhou 2010Asian Games.After a year of intensive water purification work,the 3-km stream has been transformed into the Seine of south China-the water is cleaner and businesses,including Starbucks and other cafés,have sprung up on its banks for tourists and citizens to enjoy.     

POPULATION TURNING POINT

正Latest census shows structural imbalance of population has replaced explosive growth to become China's top challenge China's population grew by less than 1 percent annually in the last decade,but it still remains the world's largest at 1.37 billion people,according to results     

Inheritors of Tibetan Thangka Art

TENPA Ramdan is a 70-year-old painter,scholar,educator and acolyte of the Mentang School of Thangka painting. In 2006,when Thangka paintings of the Mentang School appeared on the first batch to make the National Intangible Cultural Heritage List,Tenpa was designated an official inheritor of the ancient art form.     

试论妨害出入边境管理犯罪及其防治

妨害出入边境管理犯罪 ,危害国家正常的出入边境管理秩序 ,属于跨境犯罪。它不但对我国内地造成危害 ,而且也祸及港、澳、台地区 ,甚至给国际社会带来一定后果。此类犯罪产生的原因具有多样性和复杂性 ,应从宣传教育、严厉打击、海防控制、严格出入边境检查、加强队伍建设、规范出入边境中介活动、加强内地与港澳台的区际合作等方面着手 ,有效地加以防范和治理     

Lost Paradise Connected

正construction workers successfully blasted through the last section of mountain leading to Metok County, Tibet, on December 15. The explosion ended an era of isolation for the region.     

网购信任形成的动态模型研究

网络购物的信任文化不是一个静态的现象，而是一个动态的过程。已经存在的信任文化或不信任文化通过网络购物的积极或消极经验，产生增强的信任文化或不信任文化，形成新的网购信任文化背景，辐射至更大范围内的潜在网购用户，使网络购物的信任文化产生自我增强效应。     

Standing Up for Life

<正>A rehabilitation center endeavors to heal the disabled and help them live a normal life Nobody could tell the man who was walking back and forth in the lobby hall of the Deyang Rehabilitation Center for People With Disabilities has anything different from normal people,