| 查看: 720 | 回复: 3 | |||
| 本帖产生 1 个 翻译EPI ，点击这里进行查看 | |||
[交流]
求助，麻烦大家帮翻译下，感谢！
|
|||
|
Production of recombinant endotoxin neutralizing protein in Pichia pastoris and methods for its purification Production of recombinant Limulus endotoxin neutralizing protein (rENP) was attained with the GS115 methylotrophic strain ofPichia pastoris transformed with a plasmid, bearing multiple ENP gene copies.The synthetic gene for Limulus ENP was cloned into the integrative plasmid pAO815 under the control of a methanol-inducible promoter. Clones containing a single enp insert were used to construct cassettes bearing 2 and 3 tandem copies of enp. These were then integrated at the HIS locus of P. pastoris GS115 (his4).Clones were chosen for their ability to produce rENP upon methanol induction in shaker flasks, and then the 1x, 2x, and 3x-enp strains were analyzed by Southern blot for the presence of the ENP gene(s). Isolate 3x5q, containing a 3x-enp cassette, was the best producer of rENP. Under optimal conditions this strain grown in a fed-batch mode produced yields of >500mg rENP/Lwith an average of 5.46mg rENP/g DCW. Purification of rENP from the clarified broth resulted in a yield of 35% and a purity of >86%. Glycosylated rENP, the main contaminant, was removed with a concanavalin-A column and characterized. The pure rENP neutralized lipopolysaccharide and had the mass, amino-acid composition and N-terminal sequence expected from the cloned gene.2002 Elsevier Science (USA). All rights reserved. |
回复此楼» 猜你喜欢
交叉科学部支持青年基金，对三无青椒是个机会吗？
已经有4人回复
-
招博士
已经有6人回复
-
限项规定
已经有8人回复
-
国家基金申请书模板内插入图片不可调整大小?
已经有5人回复
-
国家级人才课题组招收2026年入学博士
已经有5人回复
-
Fe3O4@SiO2合成
已经有6人回复
-
青年基金C终止
已经有4人回复
-
青椒八年已不青，大家都被折磨成啥样了？
已经有7人回复
-
为什么nbs上溴 没有产物点出现呢
已经有10人回复
-
救命帖
已经有11人回复
» 抢金币啦！回帖就可以得到:
-
限广州，征女友
+2/96
-
非粮生物质能技术全国重点实验室合成生物学创新团队全球招聘博士/博士后
+1/80
-
【2026/2027 哈工大计算机类博士招生】
+1/78
-
南京林业大学特聘教授招聘博后和博士研究生
+1/75
-
广东工业大学自动化学院鲁仁全教授团队刘勇华老师招收2026年博士研究生（申请制）
+1/74
-
关于本子打包
+1/67
-
征婚
+1/60
-
华南理工大学(985、广州) 氢能源技术课题组诚招博士后（不限专业）
+1/49
-
教育部重点实验室和清华大学某国家重点实验室，联合培养硕生、博生，并长期招博士后
+1/36
-
教育部重点实验室和清华大学某国家重点实验室，联合培养硕生、博生，并长期招博士后
+1/35
-
南开大学物理学院张书辉副教授招收凝聚态物理理论方向博士生、硕士生
+1/30
-
招聘农用化学产品销售一名，须具备良好的英语口语，以便拓展海外市场。
+1/17
-
博士后, 博士招生 美国大纽约地区 NJIT
+1/11
-
美国密苏里大学堪萨斯城分校（UMKC）生物材料诚聘全奖博士
+1/11
-
山东大学集成电路学院王凌云研究员招收2026年硕士生及联合培养硕士生
+1/6
-
广东以色列理工学院博士/硕士招生-通过稀疏观测用数据驱动方法预测湍流
+1/3
-
【博士后/科研助理招聘-北京理工大学-集成电路与电子学院-国家杰青团队】
+1/2
-
华南理工大学宋波教授招收2026年博士生（二氧化碳有机、高分子和光电生物催化还原）
+1/2
-
求博导收留
+1/2
-
浙江大学 “分子智造”课题组 诚聘 博士后及科研助理
+1/1
wd519wd706(金币+20, 翻译EPI+1): 感谢你 2011-04-09 02:17:40
|
Production of recombinant endotoxin neutralizing protein in Pichia pastoris and methods for its purification 毕赤酵母中重组内毒素中和蛋白的生产及其纯化方法。 Production of recombinant Limulus endotoxin neutralizing protein (rENP) was attained with the GS115 methylotrophic strain of Pichia pastoris transformed with a plasmid, bearing multiple ENP gene copies. The synthetic gene for Limulus ENP was cloned into the integrative plasmid pAO815 under the control of a methanol-inducible promoter. 通过使用带有多种ENP基因拷贝的质粒转染GS115甲醇毕赤酵母菌株，从而获得鲎重组内毒素中和蛋白产物。鲎ENP的合成基因在甲醇诱导启动子调控下被克隆到整合质粒pAO815上。 |
2楼2011-04-08 15:20:09
|
Clones containing a single enp insert were used to construct cassettes bearing 2 and 3 tandem copies of enp. These were then integrated at the HIS locus of P. pastoris GS115 (his4). Clones were chosen for their ability to produce rENP upon methanol induction in shaker flasks, and then the 1x, 2x, and 3x-enp strains were analyzed by Southern blot for the presence of the ENP gene(s). 含有单个ENP插入物的克隆被用来构建含有2或3个连续ENP的拷贝框。然后把他们整合到毕赤酵母GS115组氨酸位点。在摇瓶中中，根据克隆在甲醇诱导下生产rENP的能力进行选择。然后用1X、2X、3XENP菌株通过Southern杂交来检测ENP基因是否存在。 Isolate 3x5q, containing a 3x-enp cassette, was the best producer of rENP. Under optimal conditions this strain grown in a fed-batch mode produced yields of >500mg rENP/Lwith an average of 5.46mg rENP/g DCW. Purification of rENP from the clarified broth resulted in a yield of 35% and a purity of >86%. 包含有3XENP框的独立3x5q（这里实在是不知道怎么翻译了，应该是个菌种品系吧），是生产rENP最好的选择。在可视条件下，这个菌株在连续补加培养模式下生长生产产量和每克5.46毫克 rENP平均干菌量想比会产生大于500毫克每升的rENP。 |
3楼2011-04-08 15:48:27
|
Glycosylated rENP, the main contaminant, was removed with a concanavalin-A column and characterized. The pure rENP neutralized lipopolysaccharide and had the mass, amino-acid composition and N-terminal sequence expected from the cloned gene.2002 Elsevier Science (USA). All rights reserved. 主要的重组形式：糖基化rENP，可以被伴刀豆球蛋白亲和色谱柱转移并标记。纯化的rENP能够中和脂多糖并且含有大量氨基酸成分和从克隆基因中所需要的N端序列。 |
4楼2011-04-08 15:54:59