Question

这句话怎么翻译？

Enzyme purification. Crude enzyme was dialyzed against 5 liters of 10 mM
glycine-NaOH buffer (pH 10.0, four buffer changes for 12 h each). The dialysate
was adjusted to pH 6.0 with 0.2 M HCl, mixed for 2 h with 500 ml of carboxymethyl cellulose equilibrate with buffer A (10 mM sodium phosphate buffer, pH
6.0), and filtered through filter paper (Whatman No. 1). The cellulose was
washed three times with 500 ml of buffer A, packed into a glass column (5 by 60
cm) and eluted by a 0;1 N NaCl gradient at a rate of 2.0 ml/min at 48C. The
active fractions (220 ml) were added to 660 ml of acetone and allowed to stand
at 48C for 18 h. The precipitates were collected by centrifugation and then
lyophilized. For further purification, gel filtration with Toyo-pearl HW 55 gel
equilibrated with 10 mM glycine-NaOH buffer (pH 10.0) was performed. The
active fractions were precipitated with acetone and then lyophilized. Sodium
dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was done according to Laemmli (15) by using a 10 to 20% gradient polyacrylamide gel and
a 4% stacking gel at 48C

Answer 1

参考翻译：酶纯化。粗酶在5升10 mM甘氨酸- naoh缓冲液中透析(pH 10.0, 4次缓冲液变化，每次12小时)。透析液的pH值调整到6.0和0.2 M盐酸混合2 h和500毫升的羧甲基纤维素与缓冲平衡(10毫米钠磷酸盐缓冲剂,pH值6.0),并通过滤纸过滤(绘画纸没有。1)。500毫升的纤维素被三次缓冲,挤进一个玻璃柱由60厘米(5)和筛选了由0;1 N氯化钠的速度梯度在48 c 2.0毫升/分钟。将活性组分(220 ml)加入660 ml丙酮中，48C静置18 h。离心收集沉淀，然后冻干。为了进一步纯化，使用以10 mM甘氨酸- naoh缓冲液(pH 10.0)平衡的Toyo-pearl HW 55凝胶过滤。活性组分经丙酮沉淀后冻干。根据Laemmli(15)，采用10 ~ 20%梯度聚丙烯酰胺凝胶和4%堆叠凝胶在48C下进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)

Answer 2

Enzyme purification. Crude enzyme was dialyzed against 5 liters of 10 mM glycine-NaOH buffer (pH 10.0, four buffer changes for 12 h each). The dialysate was adjusted to pH 6.0 with 0.2 M HCl, mixed for 2 h with 500 ml of carboxymethyl cellulose equilibrate with buffer A (10 mM sodium phosphate buffer, pH 6.0), and filtered through filter paper (Whatman No. 1). The cellulose was washed three times with 500 ml of buffer A, packed into a glass column (5 by 60 cm) and eluted by a 0;1 N NaCl gradient at a rate of 2.0 ml/min at 48C. The active fractions (220 ml) were added to 660 ml of acetone and allowed to stand at 48C for 18 h. The precipitates were collected by centrifugation and then lyophilized. For further purification, gel filtration with Toyo-pearl HW 55 gel equilibrated with 10 mM glycine-NaOH buffer (pH 10.0) was performed. The active fractions were precipitated with acetone and then lyophilized. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was done according to Laemmli (15) by using a 10 to 20% gradient polyacrylamide gel and a 4% stacking gel at 48C

酶纯化。粗酶在5升10 mM甘氨酸- naoh缓冲液中透析(pH 10.0, 4次缓冲液变化，每次12小时)。透析液的pH值调整到6.0和0.2 M盐酸混合2 h和500毫升的羧甲基纤维素与缓冲平衡(10毫米钠磷酸盐缓冲剂,pH值6.0),并通过滤纸过滤(绘画纸没有。1)。500毫升的纤维素被三次缓冲,挤进一个玻璃柱由60厘米(5)和筛选了由0;1 N氯化钠的速度梯度在48 c 2.0毫升/分钟。将活性组分(220 ml)加入660 ml丙酮中，48C静置18 h。离心收集沉淀，然后冻干。为了进一步纯化，使用以10 mM甘氨酸- naoh缓冲液(pH 10.0)平衡的Toyo-pearl HW 55凝胶过滤。活性组分经丙酮沉淀后冻干。根据Laemmli(15)，采用10 ~ 20%梯度聚丙烯酰胺凝胶和4%堆叠凝胶在48C下进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)

Answer 3

酶纯化。粗酶用5升10 mM甘氨酸NaOH缓冲液透析（pH 10.0，每种缓冲液更换4次，每次12小时）。用0.2 M HCl将透析液调节至pH 6.0，与500 ml羧甲基纤维素与缓冲液A（10 mM磷酸钠缓冲液，pH 6.0）平衡混合2 h，并通过滤纸（Whatman 1号）过滤。用500 ml缓冲液A洗涤纤维素三次，装入玻璃柱（5×60 cm）中，并在48℃下以2.0 ml/min的速率用0；1 N NaCl梯度洗脱。将活性组分（220 ml）添加到660 ml丙酮中并在48℃静置18 h。通过离心收集沉淀，然后冷冻干燥。为了进一步纯化，用东洋珍珠hw55凝胶与10mm甘氨酸NaOH缓冲液（ph10.0）进行凝胶过滤。活性组分用丙酮沉淀，然后冻干。十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）根据Laemmli（15）的要求，在48℃下用10%至20%的梯度聚丙烯酰胺凝胶和4%的堆积凝胶进行。

Answer 4

酶的净化。原酶要在5升10mm甘氨酸氢氧化钠缓冲液（pH10，4个缓冲器12小时交替用)。透析液调整到pH为6,0.2M氯化氢，和500毫升羧甲基纤维素混合，调匀到缓释液A（10mm磷酸钠缓释液，pH6），再通过过滤纸过滤（沃特曼1号）。用500毫升缓释液A清洗纤维素三遍，装进玻璃柱（5*60cm）,再用0.1氯化钠在48度下每分钟2毫升的频率洗提。活跃的分馏部分（220ml）加入到丙酮(660ml)然后保持在48度18个小时。经过离心沉淀，沉淀物被收集然后再低温冻干。为了进一步净化，用Toyo-珍珠HW55凝胶进行过滤，和10mm的甘氨酸氢氧化钠缓释液（pH值10）调匀。活跃的分馏物和丙酮沉淀下来再冻干。根据Laemmli(15)在48度环境下使用10-20%聚丙烯酰胺凝胶和4%堆胶完成十二烷基硫酸铵-聚丙烯酰胺凝胶电泳。