Ethernet over PDH技术中GFP封装模块的设计与实现

介绍Ethernet over PDH技术中GFP封装模块的设计,该设计遵循ITU-TG.7041[1]制定的GFP协议,可作为通用模块完成以太网MAC帧封装和解封装过程,应用于各种非以太网线路传输以太网数据的场合。本设计提供cHEC单bit纠错功能、将突发的以太网传输变为流畅、连续的数据流。设计采用Modelsim仿真工具完成功能仿真,通过Xilinx的XC3S1400AFT256-5芯片完成验证,并成功应用于一款以太网转多路E1芯片。     

天山雪莲△~9硬脂酰脱饱和酶基因信号肽的功能分析

为了验证天山雪莲△9硬脂酰脱饱和酶基因sikSAD信号肽的功能,本研究利用PCR法从已构建的天山雪莲cDNA文库中克隆了sikSAD的96bp信号肽序列,将其与报告基因GFP连接,成功构建了融合基因植物表达载体pCMBIA2301-sp-GFP。通过农杆菌介导的叶盘法转化烟草NC89,获得转基因植株。通过激光共聚焦显微镜观察GFP在转基因烟草叶片中的激发荧光,确定天山雪莲△9硬脂酰脱饱和酶sikSAD定位于天山雪莲的叶绿体中。     

绿色荧光蛋白研究的三个里程碑——2008年诺贝尔化学奖简介

2008年10月8日,美国科学家下村修、马丁•查尔非以及钱永健因为在绿色荧光蛋白的发现以及改造方面的突出成就而获得2008年度诺贝尔化学奖。本文拟对绿色荧光蛋白的研究历程以及应用领域做一些粗浅介绍。     

Foreign gene transfer into Chinese shrimps (Penaeus chinensis) with gene gun

Plasmids pG DNA-RZ1 with a GFP (green fluorescent protein) reporter gene and a ribozyme gene incising penaeid white spot baculovirus (WSBV) were first introduced into the fertilized eggs of Chinese shrimps by gene gun. The treated and control samples of different development stages were observed with a fluorescent microscope. The transient expression of GFP gene was high in nauplius and zoea larvae. Results from RT-PCR and PCR for adults showed that the foreign genes had been transferred into the shrimps and had expressed the corresponding proteins. This work has established a transgenic method for penaeid shrimps, which will set base for the application of genetic engineering breeding into industry.     

粪肠球菌中具有启动功能的DNA片段的捕获及鉴定

PCR扩增来自质粒pUC119的β-内酰胺酶基因(bla)作报告基因,克隆进质粒pNZ8037上的能被Nisin诱导的启动子PnisA下游,构建了粪肠球菌启动功能片段探测载体pNZ-bla.利用该探测载体从粪肠球菌染色体总DNA中克隆到了能在粪肠球菌中表达的一系列具有启动功能的DNA片段,将包含克隆片段的pNZ-P-bla6质粒电击转化粪肠球菌,培养转化子并检测其抗氨苄强度,结果表明其抗氨苄强度大于Nisin诱导的含pNZ-bla质粒的粪肠球菌.表明克隆的功能片段启动能力强于PnisA.在pNZ-P-bla6的启动功能片段下游Sau3AⅠ和EcoRⅠ之间接上绿色荧光蛋白基因作为标记,构建表达绿色荧光蛋白的质粒载体pNZ-P-gfp并导入粪肠球菌中,共聚焦显微镜下可以观察到绿色荧光蛋白的表达.     

紫花苜蓿转录因子MsDREB1基因表达产物的亚细胞定位

利用生物信息学方法对紫花苜蓿MsDREBl进行了生物信息学分析．结果表明,该序列舍有AP2典型结构域,在N端存在核定位信号．为进一步验证该基因功能,构建MsDREBl与绿色荧光蛋白（GreenFluorescentProtein,GFP）基因融合的植物表达载体pCAMBIAl302-MsDREBl,再利用基因枪将其转入洋葱表皮细胞,在共聚焦扫描显微镜下观察MsDREBl基因表达产物在洋葱表皮细胞中的亚细胞定位．结果表明,MsDREBl基因表达产物定位于细胞核中,符合DREB家族转录因子特性．     

Fluorescence Tracking of Exogenous DNA in Genetic Transformation of the Chinese Oak Silkmoth Antheraea Pernyi via Sperm-Mediated Gene Transfer

Exogenous DNA expressing green fluorescent protein（ GFP） and labeled with fluorescein isothiocyanate（ FITC） was used to transform the Chinese oak silkmoth Antheraea pernyi（ A. pernyi）via sperm-mediated gene transfer（ SMGT）. Sperms entry into the female reproductive system and eggs were observed using fluorescence microscopy. The ability of A. pernyi sperms to uptake exogenous DNA was confirmed,and transfer of the exogenous DNA was shown by GFP expression in the transgenic eggs. Our result suggested that SMGT could also be used to directly generate transgenic A. pernyi expressing functional genes of interest.     

揭示生命奥秘的荧光——2008年度诺贝尔化学奖成果简介

瑞典皇家科学院将2008年度诺贝尔化学奖授予美国日籍科学家下村修(Osamu Shimomura)、美国科学家马丁·查非(Martin Chalfie)和美籍华裔科学家钱永健(Roger Yonchien Tsien),以表彰他们在发现荧光蛋白质、改造和开发其在生物及相关领域研究的应用中做出的杰出成就。下村修首先在水晶水母(Aequorea victoria)的发光器官里发现了绿色荧光蛋白质(Green Fluorescence Protein,GFP);查非的研究揭示GFP可以在别的生物如细菌和线虫表达、发光,并可用于各种蛋白质在生物体内的示踪;钱永健则对GFP的结构和发光机理进行了深入研究,将GFP改造成各种颜色的荧光蛋白质和多种便于使用的型式,使它们在各类研究甚至非研究领域中获得广泛应用。三人专业背景和贡献方式各不相同,就像一场最终获得冠军的接力赛跑,三人都扮演了关键的角色,对于强调创新的今天都具有深刻启示作用。     

以UPS构建的酿酒酵母表达载体及GFP的表达

利用通用载体质粒融合系统UPS构建出了以GFP（绿色荧光蛋白）为报告基因的酵母表达载体，经PCR、电泳、测序等手段证明了构建的准确性，同时验证了UPS的高效性及简便性。     

Green fluorescent protein （GFP） as a vital marker for studying the interaction of Phytophthora sojae and soybean

Transgenic Phytophthora sojae strains that produce green fluorescent protein （GFP） were obtained after stable DNA integration using the Hsp70 promoter and the Ham34 terminator of Bremia lactucae. The expression of GFP during different developmental stages of P. sojae was observed using fluorescent microscopy. Based on this reporter system, the histopathologic events caused by the pathogen in soybean leaves, hypocotyls and roots were monitored. Meanwhile, the difference in resistance between different soybean cultivars against P. sojae was analyzed microscopically in roots. The results indicate that GFP can be stably expressed in zoosporangia, zoospores, cysts, hyphae and oospores of P. sojae. Using the GFP marker, the infecting pathogens in leaves, hypocotyls and roots of host could be distinctly visualized. The germ tube length of cysts germinating on the roots of resistant cultivar Nannong 8848 was longer than that on the roots of susceptible cultivar Hefeng 35. These results show for the first time that this eukaryotic reporter can be used in P. sojae as a stable and vital marker, allowing the study of genetics of this hemibiotrophic pathogen.