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用大肠杆菌同源重组获得克隆化重组腺病毒基因组 被引量:9
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作者 李文辉 王树蕙 +2 位作者 张云 刘力 public.east.cn.net 《中国生物化学与分子生物学报》 CSCD 2000年第3期346-351,共6页
利用大肠杆菌细胞内质粒间同源重组获得克隆化重组腺病毒基因组 DNA,高效构建携带有外源基因的均一重组腺病毒 .将带有狂犬病毒糖蛋白 (GP)基因和加强型 GFP(enhanced GFP,EGFP)表达盒的重组穿梭质粒 p Ad- Track- CMV/ GP与腺病毒骨架... 利用大肠杆菌细胞内质粒间同源重组获得克隆化重组腺病毒基因组 DNA,高效构建携带有外源基因的均一重组腺病毒 .将带有狂犬病毒糖蛋白 (GP)基因和加强型 GFP(enhanced GFP,EGFP)表达盒的重组穿梭质粒 p Ad- Track- CMV/ GP与腺病毒骨架载体质粒 p Ad Easy- 1一起同时电击共转化大肠杆菌 BJ51 83.在 BJ51 83细胞内 ,带有同源序列的重组穿梭质粒与骨架载体可进行同源重组 ,得到以质粒形式存在的克隆化重组腺病毒基因组 p Ad- GP’.以 p Ad- GP’为模板 ,经DNA测序确认 GP基因成功整合入此质粒中的腺病毒基因组 E1区外源基因表达盒中 .线形化的p Ad- GP’转染 2 93细胞后可得到基因组结构均一、在 E1区插入有 GP和 EGFP表达盒的重组腺病毒 ,病毒滴度可达 1× 1 0 8pfu/ ml.电镜下此重组病毒颗粒直径约为 70 nm,略呈球形 ,用荧光显微镜观察感染细胞有很强的 EGFP表达 .实验表明 :利用大肠杆菌同源重组获得克隆化的重组腺病毒基因组 DNA。 展开更多
关键词 大肠杆菌 同源重组 重组腺病毒 腺病毒载体
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人视黄醇结合蛋白在大肠杆菌中的高效表达及其活性测定 被引量:7
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作者 梁学颖 黄英武 +4 位作者 刘云 刘志红 王洪 徐琪寿 public.east.cn.net 《中国生物化学与分子生物学报》 CSCD 2000年第3期421-424,共4页
Retinol\|binding protein(RBP)is the transport protein of vitamin A and is one of the members in the family of proteins which may all bind small hydrophobic molecules.In order to study the structure\|function relations... Retinol\|binding protein(RBP)is the transport protein of vitamin A and is one of the members in the family of proteins which may all bind small hydrophobic molecules.In order to study the structure\|function relationship of RBP,the plasmid pET3a\|RBP was constructed,and RBP was expressed in E.coli Bl21(DE3)successfully.The expressed protein,with molecular weight of about 21 kD,was about 36.95 percent of the total bacteria protein as detected by SDS\|PAGE and densitometry analysis.Then the production was confirmed by probing Western blot of E.coli lysate separated on SDS\|PAGE with antisera against hRBP.In succession,the fluorescence enhancement properties and shift in absorption spectrum had also confirmed the production’s functional characteristics.It could bind ROH in ratio of 1∶1 and K d=1.5×10 -7 mol/L. 展开更多
关键词 视黄醇结合蛋白 大肠杆菌 基因表达 生物活性
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Spatial and temporal distribution of total ozone over China
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作者 Gao Qing xian, Ren Zhen hai Institute of Atmospheric Physics, Chinese Academy of Sciences, Beijing 100012, China. E mail: gaoqx@public.east.cn.net Zhang Yun gang Center of Satellite Meteorology, Beijing, China Zhang Cheng chang Beijin 《Journal of Environmental Sciences》 SCIE EI CAS CSCD 1999年第3期361-366,共6页
In this paper the grid data of total ozone mapping spectrograph (TOMS) installed on Nimbus 7 satellite (1978 to 1994) was used and the spatial and temporal distribution of total ozone over China was analyzed. The res... In this paper the grid data of total ozone mapping spectrograph (TOMS) installed on Nimbus 7 satellite (1978 to 1994) was used and the spatial and temporal distribution of total ozone over China was analyzed. The research indicates that the Qinghai Tibet Plateau destroyed the latitudinal distribution of total ozone of China and the low value closed center emerged over Qinghai Tibet Plateau. Long time change trends of seasonal total ozone of Qinghai Tibet Plateau are provided. It shows that the most obvious decrease of total ozone occurs in winter (Jan.), then in summer (Jul.), the relevant slow change occurs in autumn (Oct.) and spring (Apr.). 展开更多
关键词 total ozone TOMS Qinghai Tibet Plateau.
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