| 查看: 752 | 回复: 3 | |||
| 本帖产生 1 个 翻译EPI ，点击这里进行查看 | |||
| 当前只显示满足指定条件的回帖，点击这里查看本话题的所有回帖 | |||
[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
366求调剂
已经有9人回复
-
材料工程085601，270求调剂
已经有37人回复
-
279学硕食品专业求调剂院校
已经有18人回复
-
290调剂生物0860
已经有31人回复
-
一志愿085802 323分求调剂
已经有13人回复
-
277求调剂
已经有23人回复
-
322求调剂，08工科
已经有4人回复
-
材料工程281还有调剂机会吗
已经有30人回复
-
化学070300 求调剂
已经有16人回复
-
化工学硕294分，求导师收留
已经有12人回复
» 抢金币啦！回帖就可以得到:
-
长江大学化工学院环境学硕、专硕最后一批调剂（理学、工学均可，线上面试）
+1/88
-
山东大学杨再兴课题组长期诚聘博士后
+2/58
-
化学、化学工程、生物与医药，微生物学、生物化学与分子生物学专业调剂
+1/41
-
26申博自荐（新能源电池方向）
+1/35
-
材料与化工专业还有大量指标，欢迎大家调剂，调剂系统已经开放
+1/30
-
2026广东医科大学生物医学工程学硕、电子信息专硕接收调剂
+1/14
-
大连大学-贵州省煤炭洁净利用重点实验室联合培养研究生 化学5人+环境工程7人
+1/12
-
湖南师范大学药物分析学2026年硕士研究生调剂
+1/11
-
新出名额：欢迎药学、生物、人工智能、计算机等有班长团支书经历加入-上海
+1/10
-
350求调剂
+1/9
-
燕山大学环化学院胡老师招收硕士研究生
+1/9
-
湖南理工大学 化学、化工调剂指标充足，录取率高，欢迎第二批调剂的学生调剂
+1/9
-
陕西科技大学轻工科学与工程学院刘老师接收08专业研究生调剂
+1/8
-
中国科学技术大学-安徽大学高薪联合招聘博士后/副研究员
+1/7
-
西安工业大学光电学院接收调剂
+1/7
-
与其发展宠物经济，不如改变年轻人的婚恋观
+1/6
-
【2026考研调剂】浙江万里学院国家级青年人才团队-锂电/电催化方向招收调剂生
+1/5
-
江西科技师范大学食品安全检测课题组招收生物学/分析化学学硕调剂生各1名
+1/4
-
线上复试：河南理工大学 化工学院 接收调剂硕士研究生
+1/2
-
浙江海洋大学石油化工学院接收工科调剂（学硕、专硕均可，近期线上面试）
+1/2
|
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
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
|
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
