启动子的强度对于控制插入片段的表达水平也很重要(即,强启动子指导高表达,而较弱的启动子可以指导低/内源表达水平)。有关细菌和真核生物启动子以及研究中使用的常见启动子的更多信息,请查看Addgene的启动子参考页面(https://www.addgene.org/mol-bio-reference/promoter-background/)。 Selectable Marker:选择性标...
质粒是双链 DNA 的片段,通常携带基因并且可以独立于染色体 DNA 进行复制。虽然它们可以在古细菌和真核生物中找到,但它们在细菌中的生物学作用更为重要,它们可以通过一种水平基因转移(horizontal gene transfer )例如,接合,(conjugation),从一种细菌传递到另一种细菌,通常为宿主提供益处,例如抗生素耐药性。这种益处可能...
由于质粒多聚化是由同源重组引起的，因此缺乏重组酶蛋白 RecA 的细菌菌株的质粒扩增可以降低多聚化频率，并且在某些情况下完全阻止质粒多聚体的形成。DH5α、HB101、NEB Stable、Stbl3等大肠杆菌菌株是recA突变菌株，具有更低的质粒多聚化频率。有研究表明，与DH5α相比，菌株HB101能够产生最高的质粒单体数量。未来...
由于质粒多聚化是由同源重组引起的,因此缺乏重组酶蛋白 RecA 的细菌菌株的质粒扩增可以降低多聚化频率,并且在某些情况下完全阻止质粒多聚体的形成。DH5α、HB101、NEB Stable、Stbl3等大肠杆菌菌株是recA突变菌株,具有更低的质粒多聚化频率。有研究表明,与DH5α相比,菌株HB101能够产生最高的质粒单体数量。未来需要...
HH-CAS-101p414-TEF1p-Cas9-CYC1t(Plasmid #43802) HH-CAS-100pRPR1_gRNA_handle_RPR1t (Plasmid #49014) HH-ZW-041pYLCRISPR Cas9P35s-N HH-ZW-040pYLsgRNA-OsU6a/LacZ(Addgene 66195) HH-ZW-039pYLsgRNA-OsU3(Plasmid 66193) HH-CAS-130pAc-sgRNA-Cas9(addgene 49330) HH-CAS-129pEM-Cas9HF...
HH-CAS-101p414-TEF1p-Cas9-CYC1t(Plasmid #43802) HH-CAS-100pRPR1_gRNA_handle_RPR1t (Plasmid #49014) HH-ZW-041pYLCRISPR Cas9P35s-N HH-ZW-040pYLsgRNA-OsU6a/LacZ(Addgene 66195) HH-ZW-039pYLsgRNA-OsU3(Plasmid 66193) HH-CAS-130pAc-sgRNA-Cas9(addgene 49330) HH-CAS-129pEM-Cas9HF...
图2: 质粒DNA单体(左)与二聚体(右)图谱(图片来源:addgene blog) 1.2 质粒多聚体形成机制:同源重组 1976年,J R Bedbrook等在Cell发表文章:《Recombination between bacterial plasmids leading to the formation of plasmid multimers》,他们发现pMB9质粒DNA在rec+大肠杆菌中呈现出各种不同的大小,更大形式的pMB9...
we integrated the DNA assembly method and the software with a robotic system named Illinois Biological Foundry for Advanced Biomanufatcuring (iBioFAB) (Supplementary Fig.1) to create a nearly fully automated workflow for PlasmidMaker. As a proof of concept, we constructed 101 plasmids across six...
Plasmids to be used for the selection of stable transformants also bear the neoR gene under the control of another copy of the SV40 early promoter. Plasmids are propagated in the recA strain HB101 and prepared by a simple alkaline lysis procedure (10). After RNase treatment and polyethylene ...
1). The ColE1-like (including p15A) and pSC101 replicons accounted for 91% of the plasmids in the Addgene database. The vast majority of these plasmids belonged to the ColE1 family (86.4%), underlining the popularity of these vectors in molecular biology (Fig. 1). Surprisingly, a ...