英语翻译Cell Growth Conditions and Protein Purification.FNR wasp

英语翻译
Cell Growth Conditions and Protein Purification.FNR was
purified from an E.coli B derivative,PK22 (crp-bs990 and
Dfnr),carrying fnr under control of the inducible T7 promoter
on the plasmid pPK823 (6).For purification of the 57Fecontaining
form of FNR for Mo¨ssbauer spectroscopy,the
inoculum was prepared by centrifuging 10 ml of overnight
culture,grown in Luria broth (7),containing 0.2% glucose and
50 mgyml ampicillin,at 5,000 3 g for 5 min.The cell pellet was
then added to 1 liter of the 57Fe-enriched M9 minimal medium
(7) prepared as follows:The M9 salts solution was passed
through a Chelex 100 (200–400 mesh) column (40 g of resin
per 1 liter of medium),which eliminated naturally abundant
56Fe from the medium as shown by inductively coupled plasma
atomic emission spectroscopy.57Fe (95.9% abundance) was
added to the medium to a final concentration of 20 mM as
ferrous ethylenediammonium sulfate (brought to pH '3 with
NaOH).The following reagent grade compounds were also
added at the indicated final concentrations:0.1 mM CaCl2,1.0
mM MgSO4,0.2% casamino acid,0.2% glucose,4 mgyml
vitamin B1,and 50 mgyml ampicillin.When cultures enriched
in 57Fe or naturally abundant Fe reached an OD600 of 0.35–
0.60,FNR synthesis was induced as described (3).Following
induction of FNR synthesis,cell cultures were sparged with
argon at 4°C for 14 hr,which appeared to improve recovery of
FNR as compared with our previous results.Purification of
FNR from cell extracts was carried out under anaerobic
conditions as described (3) except that DTT was added to the
purification buffer to a final concentration of 1 mM.To
minimize any exposure to oxygen,samples were manipulated
either under a stream of copper scrubbed-argon gas or in an
anaerobic chamber (Coy),which had an atmosphere of 5%
CO2y5% H2y90% N2.
觉得多的话只翻译这两句可以不,说了不要翻译软件翻译的东西,太乱了
The M9 salts solution was passed
through a Chelex 100 (200–400 mesh) column (40 g of resin
per 1 liter of medium),which eliminated naturally abundant
56Fe from the medium as shown by inductively coupled plasma
atomic emission spectroscopy.

细胞生长条件和蛋白质纯化.FNR的是纯化大肠杆菌B衍生物,PK22（C反应蛋白,bs990和
Dfnr）,携带根据FNR的诱导T7启动子控制对质粒pPK823（6）.为了净化的57Fecontaining
FNR的形式的钼¨穆斯堡尔光谱,接种准备过夜离心10毫升文化,卢里亚肉汤成长（7）,含0.2％葡萄糖,50 mgyml氨苄青霉素,在3 5分钟5000克.沉淀的细胞然后加入1 57Fe的富升M9的基本培养基（七）编制如下：M9的盐溶液通过通过Chelex 100（200-400目）柱（40克树脂中每1公升）,其中丰富的自然淘汰从中期56Fe就表明了电感耦合等离子体原子发射光谱.57Fe的（95.9％丰度）为添加到中等至终浓度为20毫米,硫酸亚铁乙二胺（赞助商与pH值'3氢氧化钠）.下列试剂级化合物也将在指定的终浓度：0.1毫米的氯化钙,1.0毫米硫酸镁,0.2％水解酪酸,0.2％葡萄糖,4 mgyml维生素B1,50 mgyml氨苄青霉素.当文化丰富在57Fe的或自然丰富的铁OD6​​00值达到了0.35 -0.60,FNR的合成诱导描述（3）.以下FNR的合成的诱导,细胞培养与sparged氩气在4 ° C下14小时,这似乎是提高采收率的FNR的,比我们以前的结果.纯化FNR的细胞提取物是从下进行厌氧除了所述的条件数码地面电视（3）被添加到净化buffer到终浓度为1毫米.要尽量减少暴露在氧气,样品被操纵要么根据铜流擦洗,氩气或在厌氧室（大队）,其中有5％的气氛CO2y5％H2y90％的氮气.