马立克病病毒中meq基因对CEF细胞chTERT mRNA转录水平的影响

目的研究鸡马立克病病毒(MDV)meq基因对鸡胚成纤维细胞(CEF)的端粒酶催化亚单位基因(chTERT) mRNA转录水平的影响,并探讨meq基因与端粒酶活性的关系。方法构建重组载体pcDNA-meq,转染CEF细胞,利用Taq- Man探针,经实时荧光定量RT-PCR检测meq基因对CEF细胞chTERT mRNA转录水平的影响,并用TRAP法测定端粒酶活性。结果meq基因能激活CEF细胞中chTERT mRNA的转录,在48h的转录水平是72h的16倍,端粒酶活性在转染后48h是转染后72h的12倍。结论meq基因可能是MDV基因组中的主要致瘤基因,可以在体外导入正常的CEF细胞中,激活端粒酶的活性。     

拿下虚拟运营牌照 蜗牛商店注入运营商基因

正春节前,苏州蜗牛数字科技股份有限公司(简称"蜗牛")作为唯家游戏公司,获得工信部发放的虚拟运营商牌照。这将给蜗牛自研游戏平台——蜗牛移动游戏商店(简称蜗牛商店)提供一个非常有利的发展机遇,注入运营商基因的蜗牛商店有望在国内众多手游平台中异军突起。     

人工免疫优化的汽车散热器功能设计

应用公理设计原理设计车用散热器时,耦合功能的存在是一个普遍问题。国内外学者提出使用人工免疫方法求解耦合功能集。针对这些算法在度量抗体优异性时存在的不足,提出了新的度量指标：耦合功能集聚度,并给出相应公式,同时在抗体生成算法中引入基因优选的方法。经在银轮股份进行车用机油散热器设计仿真实验,表明新的算法具有较高的效率。     

表达HIV-1 Gag-Pol抗原的重组痘病毒的构建及免疫原性

目的构建稳定高效表达HIV-1Gag-Pol蛋白的重组痘病毒载体疫苗,并检测其体液免疫效果。方法将修饰型HIV-1Gag-Pol基因插入到痘病毒表达载体pSC11m2启动子P7·5下游,经与野生型痘病毒同源重组后,进行蓝白斑筛选。用筛选得到的重组病毒rM-GP转染HEK293细胞,经SDS-PAGE和Westernblot检测表达蛋白。用该重组病毒免疫BALB/c小鼠,并检测体液免疫应答。结果已筛选到稳定高效表达HIV-1Gag-Pol蛋白的重组痘病毒载体rM-GP,经Westernblot,在免疫小鼠血清中检测到抗HIV-1p24蛋白的特异性抗体。结论重组痘病毒载体rM-GP能稳定高效表达HIV-1Gag-Pol蛋白,并能引起有效的体液免疫应答。     

康宁木霉cbh2基因在毕赤酵母中的表达

为拓宽酵母菌的应用领域,构建具有分泌纤维素酶能力的酵母工程菌,利用质粒pPICZαA构建了康宁木霉纤维素酶cbh2基因的酵母表达载体,经线性化后采用电穿孔法转入毕赤氏酵母菌中.以甲醇为诱导物进行诱导,聚丙烯酰胺凝胶电泳检测结果证明该基因可成功表达.对重组蛋白进行了产酶活性测定,结果证明该重组蛋白具有纤维二糖水解活性,从而获得了一株可产生纤维素酶的酵母工程菌.     

一种新型的Tet-off慢病毒调控系统的构建及其调控作用

四环素调控的真核表达系统是真核细胞诱导表达系统的重要组成部分.当前大多数研究者采用双质粒四环素表达调控系统调控外源基因的表达.由于,双质粒四环素表达调控系统的基因表达调控严格依赖于两个单载体同时感染同一个细胞,因而限制其在体内实验中的效率.为解决该问题,研究构建了由慢病毒载体构成的单质粒四环素表达调控系统-pLV300(Tet-off).构建的载体经酶切鉴定及测序均正确.通过荧光观察及RT-PCR实验结果显示,该单质粒四环素调控的慢病毒载体体外可以高效地转移并且维持目的基因的表达,其表达调控效应高、无明显毒副作用,且调控严密.该表达调控系统为慢病毒的临床应用奠定了一定基础.     

固定化基因工程菌应用于水环境污染治理的研究进展

固定化细胞技术和基因工程技术在环境污染物治理领域是新兴技术。本文综述了近年来固定化细胞技术和基因工程技术在水环境污染治理中的研究及应用情况。通过固定化基因工程菌处理废水的一系列成功实验,表明了其在废水好氧反硝化等水环境污染治理中的广阔应用前景。     

含气脂质体的超声波介导体外基因输送的实验研究

研究不同超声频率对含气脂质体声致基因转染效率的影响.采用物理及化学法制备含气型脂质体,并比较不同磷脂配比的含气脂质体含气量差异.以不同频率超声联合化学法制备的含气脂质体转染A549细胞,比较各组报告基因的转染效率.结果表明:应用化学法制备的含气脂质体含气量显著高于物理法,脂质体含气量在卵磷脂与胆固醇配比为7∶3时达到最大值(21%);频率为45kHz的超声波转染效率显著高于其他频率对照组(p0.001),其中以含气脂质体与质粒电荷比1∶1与1∶2时为最佳.超声波联合含气脂质体可以成功介导报告基因的转染,是一种极有希望的基因输送方法.     

Expression of green fluoscrescent protein gene in Sclerotinia sclerotiorum

Protoplasts of the pathogenic plant fungus,Sclerotinia sclerotiorum,were transformed using the pPGF plasmid,which contains green fluorescent protein gene,under the control of Aspergillus nidulans regulatory sequences. The pPGF plasmid was introduced by PEG/CaCl2 treatment. Positive transformants were harvested with hygromycin B (HYG) resistance as selective marker,and then were observed with green fluorescence phenomena in response to blue light,which suggested that GFP gene was cloned into genome DNA of S. sclerotiorum. The transformants were verified mitotically stable by Southern blotting analysis and passage culturing. This study is developed as an initial step for further research into infection mechanisms of S. sclerotiorum to plants and interactions with bio-control fungus.     

Isolation and identification of a sulfate reducing bacteria and sequence analysis of its dissimilatory sulfite reductase gene

A sulfate reducing bacteria was isolated from mining sewage of Daqing Oilfield by Hungate anaerobic technology. Physiological-biochemical analysis showed that the strain could utilize polyacrylamide as sole carbon and nitrogen source. The sequence analysis of 16S rDNA illustrated that the similarity of F8 and Desulfovibrio desulfuricans (AF192153) was 99%, and the similarity sequence of dissimilatory sulfite reductase gene (DSR) cloned from the strain and Desulfovibrio desulfuricans (AF273034) was 98%. Their phylogenitic analysis was basically anastomosed, and thus temporarily named as Desulfovibrio desulfuricans F8. The DSR cloned from F8 strain was 2740 bp in length consisting of three ORF, DSRA, DSRB and DSRD as a single operon (DSRABD) regulated by the same operator. DSRA contained typical conservative box of sulfate—sulfite reducing enzyme (SiteⅠand SiteⅡ), which could bind siroheme and [Fe4S4]. DSRB retained a [Fe4S4] binding site, with an uncomplimentary structure for siroheme binding. There was no conservative box in DSRD. Sequence analysis of DSR will provide a theoretical basis for quantitative detection, metabolic pathway modification through gene engineering, and sulfate reducing bacteria (SRB) suppression.