The construction of metagenomic library from the sediment of the Zhongshan Station, Antarctica and the clone, expression and analysis of the alkaline protease ACPRO001

本文从南极中山站排污入海口（ZSS）沉积物样品中直接提取环境样品微生物总DNA，大小在48Kb以上。经过吹打调整DNA片段大小（到30-45Kb）后构建了ZScosmid宏基因组文库，获得了约6500个阳性克隆子。推算该DNA文库的总容量为228Mb，可以覆盖约10株类似E.coli菌株的总基因组。 采用蛋白酶活性筛选功能板筛选该文库我们得到了一个编号为1146的阳性克隆能够分解底物脱脂牛奶，在筛选平板上产生清晰的透明水解圈。DNA测序后分析表明该克隆外源DNA包含一个由425个氨基酸编码组成的蛋白质的完整的开放阅读框(ORF)，GC含量为50.6%。BLAST软件分析该外源DNA包含一个完...In this study, the metagenomic DNA were exrtracted and purified from a sediment sample core from ZSS. The samples ZSS was collected from the coast near the China Zhongshan Station.and the site ZSS was the sediment within the treated sewage effluent to the sea. The size of the DNA was more than 48Kb. Than the size was adjusted to 30-45Kb and was used to constract the metagenomic cosmid library. The...学位：理学硕士院系专业：生命科学学院生物医学科学系_细胞生物学学号：2172006115219

Impact of p38MAPK and RGS16 to the apoptosis and cell cycle of the glioma C6 cells

目的　探讨p38MAPK和RGS16对大鼠胶质瘤C6细胞的生物学特性的影响 .方法　利用脂质体介导法将p38MAPK和RGS16基因分别或共转染导入C6细胞中 ;用p38MAPK抑制剂SB 2 0 2 190处理处于对数生长期的转染和未转染 p38MAPK的C6细胞 ,2 4～ 36h后在倒置显微镜下观察细胞形态变化和贴壁情况 ;免疫细胞化学法检测转染前后p38MAPK和RGS16蛋白的表达情况 ;流式细胞仪检测细胞周期变化和细胞是否有凋亡发生 .结果　转染 pCMV5 p38和 /或pCMV5 RGS16质粒 36h后 30 %细胞贴壁性降低 ,突起收缩 ,细胞变圆 ;p38MAPK和RGS16蛋白均表达阳性 ;转染pCMV5 p38组出现 33.8%的凋亡峰 ,细胞周期结果显示G1期细胞百分数增加 17% ,而S期细胞百分数减少 14 % ;转染pCMV5 RGS16组无凋亡发生 ,细胞周期结果显示G1期细胞百分数减少 10 % ,而S期细胞百分数增加 14 % ;共转染pCMV5 P38和pCMV5 RGS16组未出现的凋亡峰 ,细胞周期结果显示G1期和S期细胞百分数变化与未处理组之间没有明... 【英文摘要】 AIM To study the effects of p38MAPK and RGS16 on the biological characteristics of glioma C6 cells. METHODS pCMV5 p38 and pCMV5 RGS16 were respectively or jointly transfected into C6 cells by lipofectin. SB 202190 was used to treat the transfected and nontransfected pCMV5 p38 C6 cells. The morphological and adhesive changes of the cells were observed under an inverted microscope. Expression of p38 and RGS16 was examined by immunocytochemical method both before and after the transfection. Flow Cytometr...高等学校骨干教师资助计划项目 ;; 留学归国人员科研启动基金项目 ([1 999] 747号