 |
 |
 |
 |
 |
 |
||
 |
 |
 |
 |
 |
| 6 | 1/1 | 返回列表 |
| 查看: 1286 | 回復(fù): 5 | ||
| 本帖產(chǎn)生 1 個(gè) 翻譯EPI ，點(diǎn)擊這里進(jìn)行查看 | ||
huojinlong9287銀蟲 (小有名氣)
|
[求助]
請高手潤色英文翻譯 生物學(xué)
|
|
|
為了構(gòu)建綠色熒光蛋白AcGFP1基因的原核表達(dá)載體pET32a-AcGFP1，并通過誘導(dǎo)使其在大腸桿菌中高效表達(dá)，本研究以pIRES-AcGFP1質(zhì)粒為模板，采用PCR技術(shù)特異性擴(kuò)增綠色熒光蛋白（AcGFP1）基因，并通過酶切和連接使其與原核表達(dá)載體pET32a（+）構(gòu)成重組子，經(jīng)PCR、酶切和測序鑒定后，重組質(zhì)粒轉(zhuǎn)化大腸桿菌Rosetta（DE3），用不同濃度的異丙基硫代半乳糖苷（IPTG）誘導(dǎo)表達(dá)綠色熒光蛋白，并通過15% SDS-PAGE鑒定。結(jié)果顯示重組質(zhì)粒pET32a-AcGFP1中的AcGFP1序列與Clontech公司的pIRES-AcGFP1質(zhì)粒的AcGFP1序列完全一致，說明成功構(gòu)建了含有綠色熒光蛋白AcGFP1的原核表達(dá)質(zhì)粒。pET32a-AcGFP1轉(zhuǎn)化Rosetta（DE3），經(jīng)不同濃度的IPTG誘導(dǎo)，SDS-PAGE檢測均獲得高效表達(dá)。 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. |
回復(fù)此樓» 猜你喜歡
277調(diào)劑
已經(jīng)有6人回復(fù)
-
341求調(diào)劑
已經(jīng)有7人回復(fù)
-
328求調(diào)劑，英語六級551，有科研經(jīng)歷
已經(jīng)有8人回復(fù)
-
293求調(diào)劑
已經(jīng)有12人回復(fù)
-
070300化學(xué)319求調(diào)劑
已經(jīng)有4人回復(fù)
-
268求調(diào)劑
已經(jīng)有9人回復(fù)
-
301求調(diào)劑
已經(jīng)有4人回復(fù)
-
0703化學(xué)336分求調(diào)劑
已經(jīng)有5人回復(fù)
-
301求調(diào)劑
已經(jīng)有4人回復(fù)
-
材料專碩306英一數(shù)二
已經(jīng)有9人回復(fù)
kakarote
金蟲 (文壇精英)
- 翻譯EPI: 14
- 應(yīng)助: 108 (高中生)
- 貴賓: 0.055
- 金幣: 5355.9
- 散金: 21690
- 紅花: 75
- 沙發(fā): 1
- 帖子: 11491
- 在線: 426.1小時(shí)
- 蟲號: 614324
- 注冊: 2008-09-27
- 性別: GG
- 專業(yè): 應(yīng)用光學(xué)
2樓2011-10-02 15:58:05
xiaokaizi
金蟲 (著名寫手)
- 翻譯EPI: 39
- 應(yīng)助: 6 (幼兒園)
- 金幣: 5209.5
- 散金: 2188
- 紅花: 8
- 帖子: 1415
- 在線: 176.9小時(shí)
- 蟲號: 1265018
- 注冊: 2011-04-14
- 性別: GG
- 專業(yè): 仿生材料
【答案】應(yīng)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樓2011-10-02 16:12:17
xiaokaizi
金蟲 (著名寫手)
- 翻譯EPI: 39
- 應(yīng)助: 6 (幼兒園)
- 金幣: 5209.5
- 散金: 2188
- 紅花: 8
- 帖子: 1415
- 在線: 176.9小時(shí)
- 蟲號: 1265018
- 注冊: 2011-04-14
- 性別: GG
- 專業(yè): 仿生材料
【答案】應(yīng)助回帖
huojinlong9287(金幣+50, 翻譯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樓2011-10-02 16:12:34
xiaokaizi
金蟲 (著名寫手)
- 翻譯EPI: 39
- 應(yīng)助: 6 (幼兒園)
- 金幣: 5209.5
- 散金: 2188
- 紅花: 8
- 帖子: 1415
- 在線: 176.9小時(shí)
- 蟲號: 1265018
- 注冊: 2011-04-14
- 性別: GG
- 專業(yè): 仿生材料
5樓2011-10-02 16:14:52
xiaokaizi
金蟲 (著名寫手)
- 翻譯EPI: 39
- 應(yīng)助: 6 (幼兒園)
- 金幣: 5209.5
- 散金: 2188
- 紅花: 8
- 帖子: 1415
- 在線: 176.9小時(shí)
- 蟲號: 1265018
- 注冊: 2011-04-14
- 性別: GG
- 專業(yè): 仿生材料
6樓2011-10-02 16:16:39
相關(guān)版塊跳轉(zhuǎn)
我要訂閱樓主 huojinlong9287 的主題更新
| 6 | 1/1 | 返回列表 |
| 最具人氣熱帖推薦 [查看全部] | 作者 | 回/看 | 最后發(fā)表 | |
|---|---|---|---|---|
|
[考研] 328求調(diào)劑，英語六級551，有科研經(jīng)歷 +3 | 生物工程調(diào)劑 2026-03-16 | 8/400 |
|
|---|---|---|---|---|
|
[考研] 308求調(diào)劑 +4 | 是Lupa啊 2026-03-16 | 4/200 |
|
|
|
[考研] 311求調(diào)劑 +8 | 冬十三 2026-03-15 | 8/400 |
|
|
|
[考研] 【0856】化學(xué)工程（085602）313 分，本科學(xué)科評估A類院�；瘜W(xué)工程與工藝，誠求調(diào)劑 +7 | 小劉快快上岸 2026-03-11 | 8/400 |
|
|
|
[考研] 274求調(diào)劑0856材料化工 +13 | z2839474511 2026-03-11 | 14/700 |
|
|
|
[考研] 材料工程專碩274一志愿211求調(diào)劑 +6 | 薛云鵬 2026-03-15 | 6/300 |
|
|
|
[考研] 302求調(diào)劑 +4 | 小賈同學(xué)123 2026-03-15 | 8/400 |
|
|
|
[考研] 285化工學(xué)碩求調(diào)劑（081700） +9 | 柴郡貓_ 2026-03-12 | 9/450 |
|
|
|
[考研] 材料與化工304求B區(qū)調(diào)劑 +7 | 邱gl 2026-03-11 | 8/400 |
|
|
|
[考研] 0854控制工程 359求調(diào)劑 可跨專業(yè) +3 | 626776879 2026-03-14 | 9/450 |
|
|
|
[考研] 070300化學(xué)學(xué)碩求調(diào)劑 +6 | 太想進(jìn)步了0608 2026-03-16 | 6/300 |
|
|
|
[考研] 中科院材料273求調(diào)劑 +4 | yzydy 2026-03-15 | 4/200 |
|
|
|
[基金申請]
有必要更換申報(bào)口嗎
 20+3
|
fannyamoy 2026-03-11 | 3/150 |
|
|
|
[考研] 332求調(diào)劑 +3 | zjy101327 2026-03-11 | 6/300 |
|
|
|
[考研] 290求調(diào)劑 +9 | ADT 2026-03-11 | 9/450 |
|
|
|
[考研] 求調(diào)劑 +5 | 一定有學(xué)上- 2026-03-12 | 5/250 |
|
|
|
[考研] 求b區(qū)學(xué)校調(diào)劑 +3 | 周56 2026-03-11 | 3/150 |
|
|
|
[考研] 310求調(diào)劑 +3 | 【上上簽】 2026-03-11 | 3/150 |
|
|
|
[考研] 考研調(diào)劑 +4 | 芬達(dá)46 2026-03-12 | 4/200 |
|
|
|
[考研] 314求調(diào)劑 +7 | 無懈可擊的巨人 2026-03-12 | 7/350 |
|