| 查看: 1300 | 回复: 5 | ||
| 本帖产生 1 个 翻译EPI ，点击这里进行查看 | ||
huojinlong9287银虫 (小有名气)
|
[求助]
请高手润色英文翻译 生物学
|
|
|
为了构建绿色荧光蛋白AcGFP1基因的原核表达载体pET32a-AcGFP1，并通过诱导使其在大肠杆菌中高效表达，本研究以pIRES-AcGFP1质粒为模板，采用PCR技术特异性扩增绿色荧光蛋白（AcGFP1）基因，并通过酶切和连接使其与原核表达载体pET32a（+）构成重组子，经PCR、酶切和测序鉴定后，重组质粒转化大肠杆菌Rosetta（DE3），用不同浓度的异丙基硫代半乳糖苷（IPTG）诱导表达绿色荧光蛋白，并通过15% SDS-PAGE鉴定。结果显示重组质粒pET32a-AcGFP1中的AcGFP1序列与Clontech公司的pIRES-AcGFP1质粒的AcGFP1序列完全一致，说明成功构建了含有绿色荧光蛋白AcGFP1的原核表达质粒。pET32a-AcGFP1转化Rosetta（DE3），经不同浓度的IPTG诱导，SDS-PAGE检测均获得高效表达。 The aim of this study was to construct prokaryotic expression vector pET32a-AcGFP1 and induced it high efficient expression in E. coli cell. AcGFP1 gene of green fluorescent protein was amplified from pIRES-AcGFP1 plasmid by PCR method, and was inserted into prokaryotic expression vector pET32a(+) by restriction enzyme digestion. After identifying by PCR, restriction enzyme digestion and sequencing method, AcGFP1 gene was induced with different concentrations of isopropyl-β-D-thiogalactopyranoside (IPTG) and detected on 15% SDS-PAGE. The results showed that the sequence of AcGFP1 gene in pET32a-AcGFP1 recombinant plasmid was consistent with AcGFP1 gene in pIRES-AcGFP1 plasmid from Clontech Ltd. The prokaryotic expression vector with AcGFP1 gene of green fluorescent protein was constructed successfully. All the pET32a-AcGFP1 plasmids transformed in E. coli Rosetta(DE3) were high efficient expression in SDS-PAGD after inducing with different concentrations IPTG. |
回复此楼» 猜你喜欢
080200学硕，机械工程专业277分，求带走！
已经有7人回复
-
377求调剂
已经有3人回复
-
295求调剂
已经有8人回复
-
086000生物与医药298调剂求助
已经有10人回复
-
0860 求调剂 一志愿国科大 348 分
已经有3人回复
-
材料调剂
已经有4人回复
-
本科211生物医学工程085409求调剂339分
已经有9人回复
-
296求调剂
已经有5人回复
-
385分 生物学（071000）求调剂
已经有11人回复
-
308求调剂
已经有3人回复
kakarote
金虫 (文坛精英)
- 翻译EPI: 14
- 应助: 108 (高中生)
- 贵宾: 0.055
- 金币: 5355.9
- 散金: 21690
- 红花: 75
- 沙发: 1
- 帖子: 11491
- 在线: 426.1小时
- 虫号: 614324
- 注册: 2008-09-27
- 性别: GG
- 专业: 应用光学
2楼2011-10-02 15:58:05
xiaokaizi
金虫 (著名写手)
- 翻译EPI: 39
- 应助: 6 (幼儿园)
- 金币: 5209.5
- 散金: 2188
- 红花: 8
- 帖子: 1415
- 在线: 176.9小时
- 虫号: 1265018
- 注册: 2011-04-14
- 性别: GG
- 专业: 仿生材料
【答案】应助回帖
| To construct prokaryotic expression vector pET32a-AcGFP1 and make it expresses efficiently in E. coli cell, AcGFP1 gene of green fluorescent protein was amplified from pIRES-AcGFP1 plasmid with PCR method, and it was inserted into prokaryotic expression vector pET32a(+) by restriction enzyme digestion. After identified by PCR, restriction enzyme digestion and sequencing, AcGFP1 gene was induced with different concentrations of isopropyl-β-D-thiogalactopyranoside (IPTG) and checkered on 15% SDS-PAGE. |
3楼2011-10-02 16:12:17
xiaokaizi
金虫 (著名写手)
- 翻译EPI: 39
- 应助: 6 (幼儿园)
- 金币: 5209.5
- 散金: 2188
- 红花: 8
- 帖子: 1415
- 在线: 176.9小时
- 虫号: 1265018
- 注册: 2011-04-14
- 性别: GG
- 专业: 仿生材料
【答案】应助回帖
huojinlong9287(金币+50, 翻译EPI+1): 2011-10-02 16:25:45
| The results showed that the sequence of AcGFP1 gene in pET32a-AcGFP1 recombinant plasmid was consistent with AcGFP1 gene in pIRES-AcGFP1 plasmid from Clontech Ltd, proving that the prokaryotic expression vector with AcGFP1 gene of green fluorescent protein was constructed successfully. And all of the pET32a-AcGFP1 plasmids transformed in E. coli Rosetta(DE3) were high efficient expression in SDS-PAGD after induced by different concentrations of IPTG. |
4楼2011-10-02 16:12:34
xiaokaizi
金虫 (著名写手)
- 翻译EPI: 39
- 应助: 6 (幼儿园)
- 金币: 5209.5
- 散金: 2188
- 红花: 8
- 帖子: 1415
- 在线: 176.9小时
- 虫号: 1265018
- 注册: 2011-04-14
- 性别: GG
- 专业: 仿生材料
5楼2011-10-02 16:14:52
xiaokaizi
金虫 (著名写手)
- 翻译EPI: 39
- 应助: 6 (幼儿园)
- 金币: 5209.5
- 散金: 2188
- 红花: 8
- 帖子: 1415
- 在线: 176.9小时
- 虫号: 1265018
- 注册: 2011-04-14
- 性别: GG
- 专业: 仿生材料
6楼2011-10-02 16:16:39
