24小时热门版块排行榜

返回列表

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

wd519wd706

银虫 (小有名气)

应助: 0 (幼儿园)
金币: 222.1
帖子: 96
在线: 21.4小时
虫号: 860239

[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

» 抢金币啦！回帖就可以得到:

查看全部散金贴

1楼 2011-04-06 12:58:12

已阅回复此楼关注TA 给TA发消息送TA红花 TA的回帖

相关版块跳转我要订阅楼主 wd519wd706 的主题更新