Production of selenomethionyl-derivatized proteins in baculovirus-infected insect cells

被引：27

作者：

Cronin, Ciaran N. ^{[1
]}

Lim, Kheng B. ^{[1
]}

Rogers, Joe ^{[1
]}

机构：

[1] Takeda San Diego, San Diego, CA 92121 USA

来源：

PROTEIN SCIENCE | 2007年 / 16卷 / 09期

关键词：

selenomethionine; baculovirus; insect cells; MAD phasing; multiwavelength anomalous diffraction;

D O I：

10.1110/ps.072931407

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A protocol is described for the production of both intracellularly expressed and secreted selenome-thionyl-derivatized recombinant proteins in baculovirus-infected insect cells. The method results in the production of recombinant soluble proteins with an SeMet occupancy of similar to 75% and with a recovery of similar to 20% that of native protein expression. The method is independent of the percentage methionine content of the protein and is reliable and consistent. Similar results are obtained using either Spodoptera frugiperda Sf9 or Trichoplusia ni High Five insect cells as the expression host, and when cultures are grown in either shake flasks or in Wave BioReactors.

引用

页码：2023 / 2029

页数：7

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