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北京石油化工學院2026年研究生招生接收調(diào)劑公告

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本帖產(chǎn)生 1 個翻譯EPI ，點擊這里進行查看

wo的海寶圓圓

新蟲 (小有名氣)

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[求助] 是關于細胞膜的翻譯，求幫忙，

我只能看出個大概，需要高手能給我詳細翻一下，謝謝
Cells from a 1-liter culture were harvested in the late exponential growth phase (cell density, 0.4 to 0.5 g of cells dry [weight] per liter), washed with 150 ml of 100 mM potassium phosphate buffer (pH 7.0), and finally suspended in 10 ml of this buffer.
The concentrated cell suspension was diluted with 20ml of 100 mM potassium phosphate buffer (pH 7.0) containing 10 mM MgSO4 and 150 mg of egg lysozyme (E. Merck AG,Darmstadt, Germany). The suspension was incubated for 30 min at 30°C. Subsequently, saturated K2SO4 was added to a final concentration of 0.15 M, which
resulted in lysis of the cells. Immediately thereafter, the lysed cell suspension was diluted with 70 ml of 100 mM potassium phosphate buffer (pH 7.0) containing 50 ug of RNase (Miles Laboratories, Ltd., Slough, United Kingdom) per ml (final concentration) and 50 ,ug of DNase (Miles Laboratories, Ltd.), per ml (final concentration). This solution was incubated for 20 min at 30°C, K-EDTA (pH 7.0) was added to a final concentration of 15 mM, and incubation was continued for 10 min. After the addition of MgSO4 (final concentration 20 mM), the mixture was centrifuged (30 min, 48,200 xg, 4°C: first high spin). The pellet containing membranes, cells, and cell debris was resuspended in 25 ml of 50 mM potassium phosphate buffer (pH 7.0) containing 10 mM MgSO4. Whole cells and cell debris were removed by centrifugation (70 min, 750 x g,4°C).The supernatant containing membrane vesicles was carefully decanted. Membrane vesicles were collected by centrifugation (30 min, 48,200 x g, 4C;second high spin). The bright-yellow pellet was resuspended in 50 mM potassium phosphate buffer (pH 7.0) containing 10 mM MgSO4 to a concentration of 35 mg of membrane protein per ml. Aliquots of 0.1 ml were rapidly frozen and stored in liquid nitrogen until use.

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1樓 2014-08-23 17:05:10

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ssssllllnnnn

至尊木蟲 (知名作家)

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翻譯EPI: 1690
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【答案】應助回帖

★ ★ ★ ★ ★ ★ ★ ★ ★ ★ ★
RXMCDM: 金幣+1, 多謝應助！ 2014-08-24 08:44:53
wo的海寶圓圓: 金幣+10, 翻譯EPI+1, ★★★★★最佳答案, 謝謝 2014-08-24 10:24:23

從一升對數(shù)生長期后期之培養(yǎng)細胞收集細胞（細胞密度為每升0.4至0.5克干重），用150ml 100 mM的磷酸鉀緩沖液（pH 7.0）洗滌，最后懸浮于10ml該緩沖液。

將該濃縮細胞懸液用含有10 mM MgSO4和150 mg卵溶菌酶（默克公司,達姆施塔特, 德國）的200 ml 100 mM磷酸鉀緩沖液（pH 7.0）稀釋。細胞懸浮液在30°C孵育30分鐘，加入飽和K2SO4至終濃度為0.15 M，以裂解細胞。隨即用含有每毫升50微克（終濃度）RNA酶（Miles實驗室，斯勞，英國）和每毫升50微克（終濃度）DNA酶（Miles實驗室，斯勞，英國）的70ml 100 mM磷酸鉀緩沖液（pH 7.0）. 溶液于 30°C孵育20分鐘后，加入K-EDTA（pH 7.0）至終濃度為15 mM，并繼續(xù)孵育10分鐘。加入MgSO4（終濃度為20 mM）后，混合液離心（30分鐘，48，200 x g， 4°C：第一次高速離心）。含有細胞膜、細胞、細胞碎片的沉淀懸浮于25 ml含有10 mM MgSO4的 50 mM磷酸鉀緩沖液（pH 7.0）。通過離心（70分鐘，750 x g，4°C）去除（為破碎的）全細胞以及細胞碎片。小心倒出（并收集）含有細胞膜囊泡的上清。（上清再次）離心（30 分鐘, 48,200 x g, 4°C：第二次高速離心）。將亮黃色沉淀懸浮于含有10 mM MgSO4的50 mM磷酸鉀緩沖液（pH 7.0）中，至終濃度為每毫升35 mg膜蛋白。分裝成0.1 ml到小份，快速冷凍并保存在液氮中直至使用。

贊一下

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2樓2014-08-24 05:48:27

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