| 出品公司: | ZYbscience |
|---|---|
| 载体名称: | pRI201-AN |
| 质粒类型: | 双子叶植物载体；农杆菌双元表达载体；双表达载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 克隆方法: | 限制性内切酶，多克隆位点 |
| 启动子: | 35S |
| 载体大小: | 10432 bp |
| 5' 测序引物及序列: | 35S promoter 5’CTATCCTTCGCAAGACCCTTC 3’ |
| 3' 测序引物及序列: | ATTCAGAATTGTCGANNNNNNNNNNNNNNNNN |
| 载体标签: | 无 |
| 载体抗性: | 卡那霉素 |
| 筛选标记: |
- ¥2000
- ZYbscience
- 中国/美国
- ZY3264
- 2025年07月13日
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- 详细信息
- 文献和实验
- 技术资料
- 保存条件：
-20℃低温保存
- 保质期：
三年
- 英文名：
pRI201-AN
- 库存：
20
- 供应商：
泽叶生物
载体基本信息
载体简介
pRI201-AN载体简介 The Ri Plasmid pRI201 is designed for the transformation and expression of target genes from plant cells. This series of binary vectors for plant transformation retains the backbone of the pRI101 vectors, including an alcohol dehydrogenase (ADH) gene-derived 5' untranslated region (5'-UTR) downstream of the cauliflower mosaic virus (CaMV promoter)-derived 35S promoter. Additionally, these vectors have a heat shock protein (HSP) gene-derived terminator in place of the nopaline synthase (NOS) gene-derived terminator, allowing higher target gene expression compared with the pRI101 vector series. Multigene transformation with a single vector can be achieved by integrating an expression cassette containing another gene (promoter + enhancer + gene of interest + terminator) into the vector's second multiple cloning site (MCS2) located downstream of the HSP terminator. The Ri Plasmid pRI201 binary vector series offers two types of vectors: pRI 201-AN DNA and pRI 201-ON DNA. pRI 201-AN DNA contains an Arabidopsis ADH-derived 5' UTR (AtADH 5'-UTR) and is suitable for dicotyledonous plant transformation. pRI 201-ON DNA contains a rice ADH-derived 5' UTR (OsADH 5'-UTR) and is suitable for monocotyledonous plant transformation. The pRI 201-AN DNA and pRI 201-ON DNA binary vectors for plant transformation have a mutant-type replication origin (Ri ori) from the Rhizobium rhizogenes Ri plasmid. These vectors also contain a replication origin (ColE1 ori) derived from the pUC plasmid; the ColE1 ori allows high copy number replication in E. coli. The vectors' multiple cloning site, located near the right border (RB) of T-DNA, allows stable target gene integration into the plant chromosome. pRI201-AN载体特征： Use pRI 201-AN DNA to transform dicotyledonous plant species Use pRI 201-ON DNA to transform monocotyledonous plant species Integrate multiple transgenes using a single vector High copy number (ColE1 ori) replication in E. coli pRI201-AN载体应用： Integration of multiple transgenes Dicotyledonous plant transformation Monocotyledonous plant transformation 参考文献： 1. Sugio, T. et al. (2008) J. Biosci. Bioeng. 105(3):300-302. 2. Nagaya, S. et al. (2010) Plant & Cell Physiol. 51(2):328-332. 3. Nishiguchi, R. et al. (1987) Mol. Gen. Genet. 206:1-8. 4. Satoh, J. et al. (2004) J. Biosci. and Bioeng. 98(1):1-8. 5. Sugio, T. et al. (2010) J. Biosci. Bioeng. 109(2):170-173.
载体序列
TGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTC CATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGA AACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCT CCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTG GCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAG CTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTAT CGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAAC AGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAAC TACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTC GGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTT TTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATC TTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATG AGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCA ATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCA CCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCGATCCTACAAGG TAGAATCCGCCTGAGTCGCAAGGGTGACTTCGCCTATATTGGACGACGGCGCGCAGAGGG CGACCTCTTTTTGGGTTACGATTGTAGGATTATCACTAAAACAATACATGAACATATTCA AATGGCAATCTCTCTAAGGCATTGGAAATAAATACAAATAACAGTTGGGTGGAGTTTTTC GACCTGAGGGCGTTAACCTTCTGTTAACCTAAAAGCTCTTGCCCAAACAGCAGAATCGGC GCTAATTGCCAGCGGCGGAACTTTTCCAGTTTCGCGAAAAATATCGCCACTGGCAAGGAA TGGGTTTGAGATGGCGAAGTCTGTCCTAAAAGCAGCGCCTGTAGTTGTAGGGTTGACGGC CTTGATGGAGCGTCATGCCGATGCCCTCTCGAGCCAACTTCAAGCACATCATCTTAAGGT TTTCCCGCCGCATTCCGAGAAGGGTATTCGAACATTCGGGCCATCGGAGGCGTCCAAGCT GCTCGGCGTTGGCGAGTCATATTTACGGCAGACCGCGTCTGAGATGCCAGAGTTGAATGT TAGCATGAGCCCAGGTGGCAGGCGAATGTTCTCAATTGAAGATATCCATGTGATTCGGAA GTATATGGATCAGGTCGGCCGCGGGAACCGGCGCTACCTGCCACATCGTCGAGGCGGCGA GCAGCTTCAGGTTATCTCTGTGATGAATTTCAAAGGTGGGTCGGGTAAGACCACCACCGC CGCGCATCTGGCGCAGTACCTCGCTATGCGCGGATATCGAGTCTTGGCCATTGATCTCGA TCCTCAAGCGAGCCTTTCTGCACTCTTTGGGAGCCAACCGGAGACGGACGTTGGCCCGAA CGAAACGCTCTACGGCGCTATAAGGTATGATGATGAGCAGGTGGCAATCGAACGAGTCGT CCGAGGGACTTACATTCCCGACCTCCACCTGATTCCTGGTAACCTTGAGCTGATGGAGTT TGAACACGATACGCCACGCGCGCTGATGAACCGCAAAGAGGGCGACACGCTCTTTTATGG TCGCATCAGCCAAGTAATTGAAGATATCGCGGATAACTATGACGTCGTGGTCATCGACTG CCCTCCCCAGCTTGGGTATCTCACGCTATCCGCATTGACTGCGGCGACGTCCATTCTTGT CACGGTCCATCCGCAGATGCTGGATGTGATGTCGATGAACCAGTTTCTGGCAATGACATC GAACCTTTTGCGTGAAATCGAGAATGCTGGCGCCAAGTTCAAGTTTAATTGGATGCGCTA TCTGATAACCCGTTTCGAACCGAGCGACGGACCACAGAACCAAATGGTAGGTTATCTGCG GTCGATTTTTGGCGAAAATGTCCTCAATTTTCCGATGCTTAAAACCACCGCGGTTTCGGA CGCTGGCCTGACAAACCAGACTCTATTCGAAGTGGAGCGTGGCCTGTTCACGCGCTCGAC CTATGATCGAGCCTTGGAGGCGATGAACGCCGTCAACGACGAGATCGAAACACTGATCAA AAAAGCATGGGGTAGGCCCACATGAGCCGGAAGCACATCCTTGGCGTCTCAACTGACGCC CCTGAGACGTCGCCCGCCGACAATAGGACGGCAAAGAACCGCTCCATGCCGCTCCTCGGC GTAACAAGGAAGGAGCGCGATCCGGCAACGAAGCTCACAGCGAACATTGGTAACGCACTG CGAGAGCAAAACGATCGTCTTAGCCGTGCCGAAGAGATCGAGCGGCGTCTCGCTGAAGGT CAGGCAGTGATAGAGTTGGATGCCTCGTCAATAGAACCGTCTTTCGTGCAGGATCGTATG CGAGGGGACATTGACGGGCTCCTTACTTCGATCCGGGAACAAGGACAGCAAGTCCCAATC CTTGTGCGACCGCATCCGAGCCAGCCGGGCCGATATCAGGTTGCCTTCGGCCACCGCCGG CTACGCGCCGTTTCAGAACTCGGACTTCCGGTCAGGGCGGTCGTTCGCGAACTGACGGAC GAGCAAGTGGTCGTAGCACAGGGTCAGGAAAACAATGTGCGCGAAGATCTTACCTTCATC GAAAAGGCGCGCTTCGCACATCGCCTGAACAGGCAGTTTTCTCGAGAGATTGTCATCGCC GCGATGTCGATCGACAAGAGCAATTTGTCCAAGATGCTTCTGCTCGTTGACGCCCTCCCC TCTGAACTGACCGATGCTATTGGTGCCGCTCCTGGTGTTGGACGGCCGAGTTGGCAACAA CTTGCCGAGCTGATTGAGAAAGTTTCTTCACCGGCCGACGTGGCTAAATATGCTATGTCG GAGGAAGTTCAAGCGCTGCCATCGGCAGAACGATTCAAGGCGGTGATCGCTAGTCTGAAG CCCAGTCGGGTTGCGCGTGGACTTCCCGAGGTCATGGCCACCCCAGACGGCACCAGAATT GCACAGGTGACGCAGAGCAAGGCCAAACTGGAAATCACGATTGACAGGAAGGCGACGCCC GATTTTGCGACCTTCGTGCTCGATCATGTGCCAGCGCTGTATCAAGCGTACCACGCTGAG AACCAACGGAAACGGGGAGAGTAAACCGCAAAAGAAAAGAGCCCCCTCAACGTCGCCGTC GCGGAAGCCCTTCTGTCTCTCTAGCGCGAACAGAATCGCATTTCCTCGAATCCTCGTCAA GAGTTTTTAGCGCCGTTTTGGTGAGCTGATTTCCTTTGCCTGCTGAAAGGTGAAAGATGA TGCAGACAGGAAGTGTAACGACGCCATTCGGGCGGCGGCCAATGACGCTTGCGCTTGTGC GGCGCCAGACGGCGCTGGCCGATATCAAACAAGGCAAGACAGCGGACAAGTGGAAGGTCT TTAGGGACGCGTCCGCGGCCATGGAACTACTTGGAATCCAGTCCAACAGTCTTGCCGTCC TTGATGCGCTATTGAGCTTTCACCCGGAAACGGAGTTGCGTCAGGAGGCACAGCTGATCG TCTTCCCGTCGAATGCTCAGCTTGCCCTTCGGGCGCATGGGATGGCTGGCGCGACTTTGC GTAGGCACATCGCCATGCTCGTGGAGTCAGGCTTGATCGTCCGGAAGGATAGCGCCAACG GAAAGCGTTACGCTCGTAAGGATGGCGCTGGTCAGATCGAGCGCGCGTTTGGCTTCGATT TGTCTCCGCTTCTCGCGCGGTCCGAAGAGCTAGCGATGATGGCACAGCAGGTGATGGCCG ATCGAGCAGCATTCAGGATGGCCAAAGAAAGTCTGACGATTTGCCGACGGGACGTTCGGA AGCTAATTACGGCAGCTATGGAAGAGGGAGCGGAGGGCGACTGGCAAGCTGTCGAGGAAG TCTATGTGGAACTTGTGGGTAGAATCCCACGCGCCCCGACGCTTGCTGATGTAGAGTCAA TTCTCGAAGAGATGTGGATGCTCCAGGAAGAGATAATCAACCGGTTGGAAATTAGAGACA ATTCAGAAAATAATAGCACCAATGCTGCCCAGAGCGAGCAGCACATACAGAACTCAAAAC CCGAATCCGTTAATGAACTTGAACCTCGCTCTGAAAAGGAGCAGGGCGCTAAGCCGAGTG AAATAGACCGGGCAAGGAGCGAGCCGATAAAAGCGTTCCCCCTCGGGATGATCCTGAAAG CGTGCCCGACCATTGGCAATTATGGGCCGAGCGGTGCGGTTGCTAGCTGGCGTGACCTCA TGTCGGCTGCGGTGGTGGTTCGGTCTATGCTGGGGGTCAGCCCGTCGGCTTACCAAGACG CGTGTGAGGCAATGGGACCGGAGAATGCGGCAGCAGCGATGGCGTGCATTTTGGAGCGAG CGAACTTCATCAATTCGCCTGGGGGCTATCTCCGAGATCTGACACGGCGGAGCGAACTCG GGAAGTTTTCACTTGGCCCGATGATAATGGCGCTCTTGAAGGCTAGCGGGCAGGGGACGT TGCGGTTTGGCTAGAATTAGCGAGTATGGAGCAGGATGGTCTGTGGTCAGCTGACCACAG ACCTAATAGGTTGAAAACATGAGCGTTTTTTGGATGATCGACAGACCATCCGATTCCCGG AGTACCAAGCGTGCTCTGATGGGAGCGATAACATTACTCAACAAGCACGAAGGCCCCATG CCGATCGTTGATCGTGAAGGAGAGCCTGCTCTACATGCGGCGGTATTTTGCCGGCCGAGG CATGTAGTCGCGGAGCACTGCCTATTTACTGCCCTAGGCACAAACGTTGACTCTTGGATC GAGCTGGCAGACAAAGCAATAACCCACACAGAGGACGATTAATGGCTGACGAAGAGATCC AGAATCCGCCGGACGGTACTGCTGCTGCCGAAGTTGAGCCGGCTGCTCCTAGAGGTAGAA GAGCAAAGAAAGCACCAGCCGAAACAGCCCGCACGGGATCGTTCAAATCCGTGAAGCCGA AAACCCGCGGCCTCAGCAACCGAGAAAAACTGGAGAAGATCGGTCAAATCGAAGCTCAGG TCGCTGGCGGCGCAACCTTGAAGGACGCCGTTAAGATCGTGGGTATTTCCGTTCAGACCT ATTATCAATGGAAGAGAGCTGCGGTTCAACCTGTCTCACAGAATCCGGCCGTGTCTGTTT CAGTTGACGATGAACTCGGCGAGTTCATCCAACTCGAGGAGGAAAATCGGCGGCTCAGAA AGCTAGCTTGTGGCAGGATATATTGTGGTGTAAACAAATTGACGCTTAGACAACTTAATA ACACATTGCGGACGTTTTTAATGTACTGGGGTGGTTTTTCTTTTCACCAGTGAGACGGGC AACAGCTGATTGCCCTTCACCGCCTGGCCCTGAGAGAGTTGCAGCAAGCGGTCCACGCTG GTTTGCCCCAGCAGGCGAAAATCCTGTTTGATGGTGGTTCCGAAATCGGCAAAATCCCTT ATAAATCAAAAGAATAGCCCGAGATAGGGTTGAGTGTTGTTCCAGTTTGGAACAAGAGTC CACTATTAAAGAACGTGGACTCCAACGTCAAAGGGCGAAAAACCGTCTATCAGGGCGATG GCCCAAATTCCCGATCTAGTAACATAGATGACACCGCGCGCGATAATTTATCCTAGTTTG CGCGCTATATTTTGTTTTCTATCGCGTATTAAATGTATAATTGCGGGACTCTAATCATAA AAACCCATCTCATAAATAACGTCATGCATTACATGTTAATTATTACATGCTTAACGTAAT TCAACAGAAATTATATGATAATCATCGCAAGACCGGCAACAGGATTCAATCTTAAGAAAC TTTATTGCCAAATGTTTGAACGATCGGGGCCCGATCATATTGTCGCTCAGGATCGTGGCG TTGTGCTTGTCGGCCGTTGCTGTCGTAATGATATCGGCACCTTCGACCGCCTGTTCCGCA GAGATCCCGTGGGCGAAGAACTCCAGCATGAGATCCCCGCGCTGGAGGATCATCCAGCCG GCGTCCCGGAAAACGATTCCGAAGCCCAACCTTTCATAGAAGGCGGCGGTGGAATCGAAA TCTCGTGATGGCAGGTTGGGCGTCGCTTGGTCGGTCATTTCGAACCCCAGAGTCCCGCTC AGAAGAACTCGTCAAGAAGGCGATAGAAGGCGATGCGCTGCGAATCGGGAGCGGCGATAC CGTAAAGCACGAGGAAGCGGTCAGCCCATTCGCCGCCAAGCTCTTCAGCAATATCACGGG TAGCCAACGCTATGTCCTGATAGCGGTCCGCCACACCCAGCCGGCCACAGTCGATGAATC CAGAAAAGCGGCCATTTTCCACCATGATATTCGGCAAGCAGGCATCGCCATGGGTCACGA CGAGATCATCGCCGTCGGGCATACGCGCCTTGAGCCTGGCGAACAGTTCGGCTGGCGCGA GCCCCTGATGCTCTTCGTCCAGATCATCCTGATCGACAAGACCGGCTTCCATCCGAGTAC GTGCTCGCTCGATGCGATGTTTCGCTTGGTGGTCGAATGGGCAGGTAGCCGGATCAAGCG TATGCAGCCGCCGCATTGCATCAGCCATGATGGATACTTTCTCGGCAGGAGCAAGGTGAG ATGACAGGAGATCCTGCCCCGGCACTTCGCCCAATAGCAGCCAGTCCCTTCCCGCTTCAG TGACAACGTCGAGCACAGCTGCGCAAGGAACGCCCGTCGTGGCCAGCCACGATAGCCGCG CTGCCTCGTCCTGTAGTTCATTCAGGGCACCGGACAGGTCGGTCTTGACAAAAAGAACCG GGCGCCCCTGCGCTGACAGCCGGAACACGGCGGCATCAGAGCAGCCGATTGTCTGTTGTG CCCAGTCATAGCCGAATAGCCTCTCCACCCAAGCGGCCGGAGAACCTGCGTGCAATCCAT CTTGTTCAATCATGCGAAACGATCCAGATCCGGTGCAGATTATTTGGATTGAGAGTGAAT ATGAGACTCTAATTGGATACCGAGGGGAATTTATGGAACGTCAGTGGAGCATTTTTGACA AGAAATATTTGCTAGCTGATAGTGACCTTAGGCGACTTTTGAACGCGCAATAATGGTTTC TGACGTATGTGCTTAGCTCATTAAACTCCAGAAACCCGCGGCTGAGTGGCTCCTTCAACG TTGCGGTTCTGTCAGTTCCAAACGTAAAACGGCTTGTCCCGCGTCATCGGCGGGGGTCAT AACGTGACTCCCTTAATTCTCCGCTCATGATCCTGGCACGACAGGTTTCCCGACTGGAAA GCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCT TTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCAC ACAGGAAACAGCTATGACCATGATTACGAATTCCCGGGATCCAGCGGCCGCAGGTACCAA TTCCTTATCTTTAATCATATTCCATAGTCCATACCATAGCACATACAGTAGTTATATGCG AAGAGATCCAACAAAACATTCACAATGGATTATAGAAACATTTGTTTATTCATTATAATG AGATCTTACATTCATTTAATATTAGAAAAAGCCACAAATTCATAACACAACAAGCCAAGA AAAAAACACAAACTTAAGCACACAAGCTAGCTTTTTATTTGACACACCAAATATTTCATC TTCATCTTCATAAGAGCTCGGTAGCAATTCCCGAGGCTGTAGCCGACGATGGTGCGCCAG GAGAGTTGTTGATTCAGAATTGTCGACCCGGTAGTAACTTGCTAATCGACGGGCATATGT ATCAACAGTGAAGAACTTGCTTTTGATCTTTTGTTAGTTTTGTGTGATTGTGATGTATCT AGAGTCCCCCGTGTTCTCTCCAAATGAAATGAACTTCCTTATATAGAGGAAGGGTCTTGC GAAGGATAGTGGGATTGTGCGTCATCCCTTACGTCAGTGGAGATATCACATCAATCCACT TGCTTTGAAGACGTGGTTGGAACGTCTTCTTTTTCCACGATGCTCCTCGTGGGTGGGGGT CCATCTTTGGGACCACTGTCGGCAGAGGCATCTTCAACGATGGCCTTTCCTTTATCGCAA TGATGGCATTTGTAGGAGCCACCTTCCTTTTCCACTATCTTCACAATAAAGTGACAGATA GCTGGGCAATGGAATCCGAGGAGGTTTCCGGATATTACCCTTTGTTGAAAAGTCTCAATT GCCCTTTGGTCTTCTGAGACTGTATCTTTGATATTTTTGGAGTAGACAAGTGTGTCGTGC TCCACCATGTTGACGAAGATTTTCTTCTTGTCATTGAGTCGTAAGAGACTCTGTATGAAC TGTTCGCCAGTCTTTACGGCGAGTTCTGTTAGGTCCTCTATTTGAATCTTTGACTCCATG GCCTTTGATTCAGTGGGAACTACCTTTTTAGAGACTCCAATCTCTATTACTTGCCTTGGT TTGTGAAGCAAGCCTTGAATCGTCCATACTGGAATAGTACTTCTGATCTTGAGAAATATA TCTTTCTCTGTGTTCTTGATGCAGTTAGTCCTGAATCTTTTGACTGCATCTTTAACCTTC TTGGGAAGGTATTTGATTTCCTGGAGATTATTGCTCGGGTAGATCGTCTTGATGAGACCT GCTGCGTAAGCCTCTCTAACCATCTGTGGGTTAGCATTCTTTCTGAAATTGAAAAGGCTA ATCTGGGGACCTGCAGGCATGCAAGCTTGGCACTGGCCGTCGTTTTACAACGTCGTGACT GGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGGA TCAGATTGTCGTTTCCCGCCTTCAGTTTAAACTATCAGTGTTTGACAGGATATATTGGCG GGTAAACCTAAGAGAAAAGAGCGTTTATTAGAATAATCGGATATTTAAAAGGGCGTGAAA AGGTTTATCCGTTCGTCCATTTGTATGTGACTAAAACAATTCATCCAGTAAAATATAATA TTTTATTTTCTCCCAATCAGGCTTGATCCCCAGTAAGTCAAAAAATAGCTCGACATACTG TTCTTCCCCGATATCCTCCCTGATCGACCGGACGCAGAAGGCAATGTCATACCACTTGTC CGCCCTGCCGCTTCTCCCAAGATCAATAAAGCCACTTACTTTGCCATCTTTCACAAAGAT GTTGCTGTCTCCCAGGTCGCCGTGGGAAAAGACAAGTTCCTCTTCGGGCTTTTCCGTCTT TAAAAAATCATACAGCTCGCGCGGATCTTTAAATGGAGTGTCTTCTTCCCAGTTTTCGCA ATCCACATCGGCCAGATCGTTATTCAGTAAGTAATCCAATTCGGCTAAGCGGCTGTCTAA GCTATTCGTATAGGGACAATCCGATATGTCGATGGAGTGAAAGAGCCTGATGCACTCCGC ATACAGCTCGATAATCTTTTCAGGGCTTTGTTCATCTTCATACTCTTCCGAGCAAAGGAC GCCATCGGCCTCACTCATGAGCAGATTGCTCCAGCCATCATGCCGTTCAAAGTGCAGGAC CTTTGGAACAGGCAGCTTTCCTTCCAGCCATAGCATCATGTCCTTTTCCCGTTCCACATC ATAGGTGGTCCCTTTATACCGGCTGTCCGTCATTTTTAAATATAGGTTTTCATTTTCTCC CACCAGCTTATATACCTTAGCAGGAGACATTCCTTCCGTATCTTTTACGCAGCGGTATTT TTCGATCAGTTTTTTCAATTCCGGTGATATTCTCATTTTAGCCATTTATTATTTCCTTCC TCTTTTCTACAGTATTTAAAGATACCCCAAGAAGCTAATTATAACAAGACGAACTCCAAT TCACTGTTCCTTGCATTCTAAAACCTTAAATACCAGAAAACAGCTTTTTCAAAGTTGTTT TCAAAGTTGGCGTATAACATAGTATCGACGGAGCCGATTTTGAAACCACAATTATGGGTG ATGCTGCCAACTTACTGATTTAGTGTATGATGGTGTTTTTGAGGTGCTCCAGTGGCTTCT GTTTCTATCAGCTGTCCCTCCTGTTCAGCTACTGACGGGGTGGTGCGTAACGGCAAAAGC ACCGCCGGACATCAGCGCTATCTCTGCTCTCACTGCCGTAAAACATGGCAAC
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zwmc 很多文章使用这种方法来表达miRNA, 而且我在论坛上看到很多坛友也比较推崇这个方法。但是我最近遇到了问题：我想要克隆表达的miRNA具有5p和3p两种剪切形式，理论上我把包含该pri-miRNA的基因组片段到表达载体在细胞里表达之后，这种情况下我表达出来的miRNA是兼有5p 3p两种形式？或者是只有一种呢？ 很困惑，请大家不吝赐教！ 吴佰霖 1. 查文献，看这个miRNA的5p和3p是否都有表达
类似物的瞬时表达方式。miRNA的过表达载体中有插入成熟miRNA、pre-miRNA及pri-miRNA序列三种形式。其中pri-miRNA（含侧翼序列）的方式由于保留了每个microRNA独自的原始天然双臂结构，效果更好，是应用最为广泛的方法。microRNA过表达系统中有CMV启动子（II型启动子）和H1启动子（III型启动子）两种方式供选择。III型启动子（H1、U6等）具有表达效率高、有效的确定终止位置等优点，成为首选。II型启动子适用于pri-miRNA序列中有连续的5个T-导致III
【求助】请问是否有带GFP的表达载体可以用了表达microRNA
。 geneyouth 用pEGFP -C3载体 就可以， pri-miR位于GFP的3‘ UTR区域 xshdao geneyouth用过pEGFP -C3载体表达pri-miR否，敬请传授经验！谢谢！ geneyouth 恩 其实过表达miRNA并没有那么复杂 直接连接入cmv 或 u6启动子后面 过表达miRNA前提就可以了。 你 多看看别人的文章 记得PNAS上有
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pRI201-AN载体
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