| 查看: 1281 | 回复: 1 | |||
| 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]
英文精细润色 生物学 （已附中文）
|
||
|
润色英文翻译 请不要灌水 高手优先 为了构建猪SRY基因的原核表达载体pET-32a(+)-SRY，并通过诱导使其在大肠杆菌中获得高效表达。本研究采用添加限制性内切酶位点的引物特异性扩增SRY基因，连入pMD19-T simple载体，转化入大肠杆菌DH5α，克隆后提取pMD19-T-SRY阳性重组质粒，使用相同的内切酶同时对pMD19-T-SRY质粒和原核表达pET-32a（+）载体进行酶切，连接后使SRY基因定向克隆到pET-32a(+)表达载体中。经PCR、酶切和测序鉴定后，重组质粒转化大肠杆菌感受态DH5α，提取质粒后再次转化大肠杆菌Rosetta（DE3），用不同浓度的异丙基硫代半乳糖苷（IPTG）诱导表达，并通过15% SDS-PAGE鉴定。结果显示，不同浓度IPTG诱导的SRY基因均在大肠杆菌中进行了高效特异性融合表达。 To construct prokaryotic expression vector pET-32a(+)-SRY of pig and induced it expresses efficiently in E. coli cell, SRY gene was amplified by using primer with restriction enzymes, and it was inserted into pMD19-T simple vector and transferred into the bacterium DH5α for replication. The pMD19-T-SRY recombinant plasmids were extracted, then pMD19-T-SRY and pET-32a（+）were digested with same restriction enzymes. By this directed cloning technique, the SRY gene was inserted into pET-32a（+）expression plasmid. After identified by PCR, restriction enzyme digestion and sequencing, The pMD19-T-SRY recombinant plasmids were transformed into competent cell DH5α. the pMD19-T-SRY plasmids were extracted and transformed to competent cell Rosetta（DE3）. pMD19-T-SRY was induced with different concentrations of isopropyl-β-D-thiogalactopyranoside (IPTG) and detected on 15% SDS-PAGE. The results showed that the fusion protein were specifically high efficient expression in SDS-PAGE after induced by different concentrations of IPTG. |
回复此楼» 猜你喜欢
考研调剂
已经有4人回复
-
285求调剂
已经有6人回复
-
一志愿北京化工大学070300 学硕336求调剂
已经有7人回复
-
求调剂
已经有6人回复
-
275求调剂
已经有4人回复
-
318求调剂
已经有6人回复
-
305分求调剂（食品工程）
已经有5人回复
-
求调剂院校信息
已经有5人回复
-
一志愿070300浙大化学358分，求调剂！
已经有3人回复
-
一志愿东华大学化学070300，求调剂
已经有5人回复
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
★ ★
爱与雨下(金币+2): ！~ 2011-10-09 20:29:47
huojinlong9287(金币+100, 翻译EPI+1): 2011-10-10 09:59:00
爱与雨下(金币+2): ！~ 2011-10-09 20:29:47
huojinlong9287(金币+100, 翻译EPI+1): 2011-10-10 09:59:00
|
谈不上高手，只是相互交流学习一下。注意有几处修改的地方（上传时标记被抹去了）。仅供参考，， To construct pig’s prokaryotic expression vector pET-32a(+)-SRY and induce it to express efficiently in E. coli cell, SRY gene was amplified by (using) primer with restriction enzymes, and it was inserted into pMD19-T simple vector and transferred into the bacterium DH5α .After replication the pMD19-T-SRY recombinant plasmids were extracted, then pMD19-T-SRY and pET-32a（+）were digested by the same restriction enzymes at the same time. By this directed cloning technique, the SRY gene was inserted into pET-32a（+）expression plasmid. After PCR identifying, restriction enzyme digestion and sequencing testing, the pMD19-T-SRY recombinant plasmids were transformed into competent cell DH5α. The pMD19-T-SRY plasmids were extracted and transformed to competent cell Rosetta(DE3). pMD19-T-SRY was induced with different concentrations of isopropyl-β-D-thiogalactopyranoside (IPTG) and identified on 15% SDS-PAGE. The results showed that the fusion protein had high efficient expression in SDS-PAGE when induced by different concentrations of IPTG. |
2楼2011-10-09 20:19:54
