■ 基本信息
啟動(dòng)子: | H1 |
復(fù)制子: | pUC |
終止子: | SV40 poly(A) signal，bGH poly(A) signal |
質(zhì)粒分類(lèi): | 病毒系列，慢病毒干擾載體 |
質(zhì)粒大小: | 11085bp |
原核抗性: | Amp |
克隆菌株: | Stbl3 |
培養(yǎng)條件: | 37度 |
表達(dá)宿主: | 哺乳細(xì)胞 |
誘導(dǎo)方式: | 無(wú)須誘導(dǎo)，瞬時(shí)表達(dá) |
5'測(cè)序引物: | 根據(jù)序列設(shè)計(jì)引物 |
3'測(cè)序引物: | WPRE-R：CATAGCGTAAAAGGAGCAACA |
■ 質(zhì)粒屬性
質(zhì)粒宿主: | 哺乳細(xì)胞,慢病毒 |
質(zhì)粒用途: | 基因沉默 |
片段類(lèi)型: | shRNA |
片段物種: |
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原核抗性: | Amp |
真核抗性: |
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熒光標(biāo)記: | 綠色
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■ 質(zhì)粒簡(jiǎn)介
pLVTHM allows for direct cloning of shRNAs into the lentiviral vectors and replaces the old pLVTH system from Trono lab. pLVTHM is similar to pLVTH, but contains a 3bp substitution that generates a unique MluI site for direct cloning of an shRNA into MluI-ClaI. Please note that there is an additional ClaI site in this vector that is blocked by Dam methylation . The plasmid needs to be grown in a Dam+ bacterial strain in order to use ClaI for cloning. Note that ClaI has lower salt concentration requirements than MluI. One way to handle this is to digest with ClaI for 45 minutes, followed by addition of diluted MluI buffer and MluI, then incubation for 90 more minutes. Be sure not to use a large amount of vector (1.7 ug to 2.5 ug is known to work). Also, if your shRNA is already in pSUPER (or another plasmid under the control of the PolIII promoter) you may use the EcoRI-ClaI sites to replace the H1 promoter in pLVTHM with the H1-shRNA cassette from pSUPER (or another plasmid).
■ 質(zhì)粒圖譜
■ 質(zhì)粒序列
質(zhì)粒序列請(qǐng)下載：
ZK822pLVTHM慢病毒干擾質(zhì)粒.txt
