Plasmid and Phagemid cloning vector - Selectable vs. Screening marker
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- Опубликовано: 24 июл 2024
- References/Resources: / the_crux
This video discusses the general biology of cloning vectors focusing on plasmids and phagemids. We discuss how plasmids replicate, their general properties, diversity, origins, and many other important details. This will also help you understand the evolution of cloning vectors from a standard Plasmid to Phagemid. This video also provides you with a conceptual framework of selectable and screening markers, discussed with examples.
Jump to your favorite section:
00:00 Outline
00:59 Natural plasmids
04:26 Plasmid size
06:26 ColE1 plasmid replication
10:40 pBR322 plasmid
12:36 pUC18/19 plasmid
14:23 Phagemid (pBlueScript)
17:26 Selectable markers
20:08 Screening markers
23:32 Blue-White screening
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Erratum: 11:13 - The origin is taken from MB1 plasmid and NOT from ColE1. Note that ColE1 and MB1 are both ori-type plasmids so their "ori" work identically (as explained at 4:36).
This series is absolutely 100% helpful especially for Biotechnologists
Keep uploading quality content like these!!! 👍
Please upload on PCR in detail mechanism and it's different types
Thanks for the lecture 😊
20:00 One can use bacterial strain such as DB3.1, which contains a mutant version of DNA gyrase (gyrA462) that is resistant to the toxic effects of CcdB. Cheers👍
Perfect 🙂
@@theCrux thanks. Your videos are fantastic and keep making more. Best wishes.
Wait I'm confused how can you tell which bacteria have disrupted plasmids if they are all immune to it's active effects? Or do you mean propagation outside of synthetic lethality? Also dumb questions what does disrupted mean for a plasmid. Also what is a starting plasmid and why is cddb toxic for that even after it is disrupted?
Hey can you please make more videos like on pET vectors and Bacterial expression system
25:53
Isn't it like this that during Alpha complementation, the Lacz alpha only produces 2 subunits of beta galactosidase and the remaining 2 omega subunits comes from the mutated E.coli strain (Delta M15) which also has a defective lacz gene, and due to complementation between these 2 (alpha and omega) makes it a functional beta galactosidase giving blue colour
And on the contrast, if our gene of interest disrupts lacz alpha gene, then only omega subunit is produced which is non functional and we get white colonies...
Yep, those details sound about right.
@@theCrux fairs...
Thank you, your videos helped so much...
Can you post a video for bacteriophages as vectors both M13 and lambda phage and how we clone our gene of interest in this system
The video for lambda phage is here ruclips.net/video/SHE0lpMiitA/видео.html