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theCrux
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Добавлен 14 дек 2018
Rigorously explaining and illustrating biological concepts.
If you would like to access resources and reading material used to make these videos or invest in the growth of the channel, please check out the Patreon page: www.patreon.com/the_Crux
If you would like to access resources and reading material used to make these videos or invest in the growth of the channel, please check out the Patreon page: www.patreon.com/the_Crux
Protein Expression Vectors - Expression Host and Recombinant Proteins [Part 1]
References/Resources: www.patreon.com/the_Crux
All videos on Genetic Engineering: ruclips.net/p/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf
Protein Expression vectors are used to make protein from the template DNA. In this video, we discuss some of the common applications of protein expression vectors and the complications involved with protein expression when changing the host i.e. heterologous protein expression. We compare prokaryotic and eukaryotic expression systems. Finally, we explore the molecular components for a protein expression vector and the extra engineering that goes with it.
Jump to your favorite section:
00:00 Goals
00:36 Application
01:45 Expectation
03:03 Host Comparison
05:30 Host Li...
All videos on Genetic Engineering: ruclips.net/p/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf
Protein Expression vectors are used to make protein from the template DNA. In this video, we discuss some of the common applications of protein expression vectors and the complications involved with protein expression when changing the host i.e. heterologous protein expression. We compare prokaryotic and eukaryotic expression systems. Finally, we explore the molecular components for a protein expression vector and the extra engineering that goes with it.
Jump to your favorite section:
00:00 Goals
00:36 Application
01:45 Expectation
03:03 Host Comparison
05:30 Host Li...
Просмотров: 111
Видео
RNA Expression Vectors - Design, Applications, and Examples (In vitro Transcription)
Просмотров 147День назад
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: ruclips.net/p/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf RNA Expression vectors (different from protein expression vectors) are used for the sole purpose of making RNA from a template DNA. In this video, we discuss some of the common applications of RNA expression vectors and their rules/principles of design. Finally, we ...
Golden Gate Cloning or assembly - Type IIS (BsaI) restriction enzyme based cloning
Просмотров 6522 месяца назад
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: ruclips.net/p/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf Golden Gate Cloning is a restriction enzyme based cloning that uses type IIS restriction enzymes. It is a powerful way to clone large numbers of inserts in a single reaction without the need of using multiple enzymes. This video discusses the basis of golden gate as...
Gateway Cloning: Simple and Multisite Gateway Cloning - LR and BP Cloning
Просмотров 1,1 тыс.4 месяца назад
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: ruclips.net/p/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf Gateway Cloning is a popular recombination based cloning method. It does not rely on restriction enzymes but instead uses integrases to recombine att sites between a final and donor vector. This causes an exchange of DNA segments that inserts the desired DNA into th...
TOPO Cloning - TOPO-Blunt, TOPO-TA, TOPO-directional
Просмотров 2,8 тыс.7 месяцев назад
References/Resources: www.patreon.com/the_Crux All videos on Genetic Engineering: ruclips.net/p/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf TOPO Cloning (Topoisomerase-based cloning) is a commonly used method to clone PCR amplicons. TOPO Cloning bypasses the need of restriction enzymes and ligases. In this video, we discuss the three main types of TOPO-cloning systems - TOPO Blunt, TOPO TA, and TOPO dir...
Cloning with Restriction Enzymes (5 Levels) - Traditional Molecular Cloning
Просмотров 1 тыс.8 месяцев назад
References/Resources: www.patreon.com/the_Crux Playlist - Genetic Engineering: ruclips.net/p/PL0Ymnd-zt4Ij9qp5ziU0TkyF42LUcgHQf Molecular Cloning with restriction enzymes can be very easy or very challenging. This video discusses the fundamental considerations of traditional restriction enzyme based molecular cloning. We discuss sticky and blunt ends, the process of blunting, and how all of thi...
Plant cloning vectors - Disarmed Ti plasmid, Cointegrate vectors, Binary vectors - T DNA transfer
Просмотров 3,2 тыс.9 месяцев назад
References/Resources: www.patreon.com/the_Crux This video discusses the plant cloning vectors focusing on Agrobacterium Tumefaciens based vectors and its dependency on the natural infection mechanism. This involves understanding Ti plasmids, disarmed Ti vectors, cointegrate vectors, and binary vectors. The process of cloning in plants uses conjugative transfer of T DNA through conjugation betwe...
Yeast cloning vectors - YIp, YEp, YRp, YCp, YACs - Shuttle plasmid vectors
Просмотров 4,1 тыс.9 месяцев назад
References/Resources: www.patreon.com/the_Crux This video discusses the general biology of yeast cloning vectors. There are 5 classic vectors - Integrating, Episomal, Replicating, Centromeric, and Artificial Chromosomes - as well as the naturally occurring Yeast 2 mu plasmid. Yeast vectors rely on coli-based vectors like pBR322, pUC19, or Bluescripts. Typically, yeast cloning vectors tend to be...
P1 Phage and PAC cloning vector - Pac sites, Cre-loxP, and SacB lethality
Просмотров 1,5 тыс.9 месяцев назад
References/Resources: www.patreon.com/the_Crux This video discusses the general biology of P1 bacteriophage based cloning vectors (pNS582 and pAd10SacBII) and P1 derived artificial chromosomes (PACs - pCYPAC1). The discussion walks you through the development of the P1 system with its use of Pac site and Cre-loxP recombination, coupled with SacB driven synthetic lethality. The main focus of the...
F plasmid, BACs, Fosmid cloning vector - Fosmids are small BACs (Bacterial Artificial Chromosomes)
Просмотров 1,2 тыс.10 месяцев назад
References/Resources: www.patreon.com/the_Crux This video discusses the general biology of F plasmids, Bacterial Artificial Chromosomes, BACs, and Fosmids. We start the origin of high capacity cloning vectors with the F plasmids and the development of mini F vectors. This was the inspiration for BACs which went through a few iterations involving retrofits and screening markers. Fosmids were dev...
Lambda Phage and Cosmid cloning vector - spi phenotype and in vitro packaged lambda virus
Просмотров 4,8 тыс.10 месяцев назад
References/Resources: www.patreon.com/the_Crux This video discusses the general biology of lambda bacteriophage and cosmid cloning vectors. There are two types - insertional and replacement - of vectors derived from lambda bacteriophage. This requires the use of spi phenotype since bacteriophage vectors have no selection or screening markers. From the bacteriophage vector, cosmids are created, ...
Plasmid and Phagemid cloning vector - Selectable vs. Screening marker
Просмотров 3,1 тыс.10 месяцев назад
References/Resources: www.patreon.com/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 ...
Dam and Dcm methylation (restriction independent methyl-transferases) affects restriction enzymes
Просмотров 1,5 тыс.10 месяцев назад
References/Resources: www.patreon.com/the_Crux This video discusses restriction independent methyl-transferases like dam and dcm methylase - their biological function and how they affect restriction enzyme behavior. Apart from Dam and Dcm methyl-transferases, we discuss EcoK1, mcrABC and mrr endonuclease as well. All this is done to recognize the importance of understanding DNA methylases and t...
Restriction Enzymes in Molecular Cloning
Просмотров 1,4 тыс.10 месяцев назад
References/Resources: www.patreon.com/the_Crux This is an introduction to restriction enzymes. We discuss the importance of restriction and modification systems in molecular immunity and how this property is exploited for use in molecular cloning. Different types of restriction enzymes are discussed with specific emphasis on type 2 systems - specifically 2P and 2S systems. From this discussion,...
Genetic Engineering - Classic vs Modern methods
Просмотров 2,5 тыс.11 месяцев назад
References/Resources: www.patreon.com/the_Crux This is an introduction to various methods used in Genetic engineering. Depending on the timescale of discovery and inventions, genetic engineering can be split into classic and modern genetic engineering. Classic methods deal with gene-level manipulation where the DNA is extracted from the cell. Modern methods deal with genome-level manipulation a...
RNA Polymerase III Transcription - Promoters, TFs, Initiation, Elongation, and Termination steps
Просмотров 3,3 тыс.11 месяцев назад
RNA Polymerase III Transcription - Promoters, TFs, Initiation, Elongation, and Termination steps
Prokaryotic Transcription Factors in Elongation and Termination (GreA, GreB, Mfd, NusA)
Просмотров 2 тыс.Год назад
Prokaryotic Transcription Factors in Elongation and Termination (GreA, GreB, Mfd, NusA)
Rolling Circle DNA Replication and Amplification - Plasmids and Bacteriophages (M13 + PhiX174)
Просмотров 9 тыс.Год назад
Rolling Circle DNA Replication and Amplification - Plasmids and Bacteriophages (M13 PhiX174)
Mitochondrial DNA (mtDNA) Replication - D loop Replication (deep-dive video)
Просмотров 11 тыс.Год назад
Mitochondrial DNA (mtDNA) Replication - D loop Replication (deep-dive video)
Telomere DNA Replication - Chromosome end replication problem in eukaryotes
Просмотров 4,4 тыс.Год назад
Telomere DNA Replication - Chromosome end replication problem in eukaryotes
Epigenetic Replication during DNA Replication - Replicating histones and DNA methylation
Просмотров 1,5 тыс.Год назад
Epigenetic Replication during DNA Replication - Replicating histones and DNA methylation
DNA Replication (Eukaryote) TERMINATION - Non-specific vs Site-specific Termination Mechanism
Просмотров 2,3 тыс.Год назад
DNA Replication (Eukaryote) TERMINATION - Non-specific vs Site-specific Termination Mechanism
DNA Replication (Eukaryote) ELONGATION - Polymerase Switching + different types of DNA polymerase
Просмотров 4,7 тыс.Год назад
DNA Replication (Eukaryote) ELONGATION - Polymerase Switching different types of DNA polymerase
DNA Replication (Eukaryote) INITIATION - Origin Firing - CMG Helicase forms the Replisome
Просмотров 2,2 тыс.Год назад
DNA Replication (Eukaryote) INITIATION - Origin Firing - CMG Helicase forms the Replisome
DNA Replication (Eukaryote) INITIATION - Origin Licensing - cdt1 recruits Helicase Double Hexamer
Просмотров 2,8 тыс.Год назад
DNA Replication (Eukaryote) INITIATION - Origin Licensing - cdt1 recruits Helicase Double Hexamer
DNA Replication (Eukaryote) INITIATION - Origin Recognition by ORCs and cdc6
Просмотров 3,7 тыс.Год назад
DNA Replication (Eukaryote) INITIATION - Origin Recognition by ORCs and cdc6
DNA Replication (Prokaryote) TERMINATION - Ter sites and Tus protein together stop DNA replication
Просмотров 4,8 тыс.Год назад
DNA Replication (Prokaryote) TERMINATION - Ter sites and Tus protein together stop DNA replication
DNA Replication ELONGATION - DNA Synthesis Biochemistry
Просмотров 2,4 тыс.Год назад
DNA Replication ELONGATION - DNA Synthesis Biochemistry
DNA Replication (Prokaryote) ELONGATION - Trombone Model of Elongation + Okazaki Fragment Synthesis
Просмотров 10 тыс.2 года назад
DNA Replication (Prokaryote) ELONGATION - Trombone Model of Elongation Okazaki Fragment Synthesis
The problem is : how long does the amount of injected mRNA continue producing SP ? An overproduction of SP ( the antigen) will exhaust the cells producing the antibody: and that is what we suspect in patients with "long covid": a massive reduction of antibody producing cells and consequently of the antibody = no cellular and no serologic immune deffences anymore- which would explain the explosion of turbocancers ! HOW to STOP mRNA's overproduction of the antigen which infiltrates tissues and creates deposits ( like in amyloidosis and other thesauropathies) Has the concentration of SP been measured and controlled in long covid patients ? I must suppose : not F de Clari MD 9.9.24
Awesome videos. Finally someone going into detail. I hope this will help me to design my first plasmids. Thank you 🙏.
i never leave youtube comments but your videos are too incredible to not thank you for making these. seriously some of the best details and explanations i have seen. would it be possible to request a video on the RNA interference pathway if you have a chance?
Glad to hear that the content is useful :) And yes, RNAi will be part of this "Genetic Engineering" series. I hope to have that video up by the end of this year.
your explanation is so excellent + illustrations makes it easy to understand!! thank you so much. do u perhaps cover topics in genetics? if so could we request a video on chromosome structure chemistry(detailed )+ linkages and recombination?
Thanks for the suggestion. I do expect to cover Genetics and chromosome biology at some point. Currently caught up in videos on Genetic Engineering :)
@@theCrux thank you so much. will be looking forward to it!
saving lives!!! thank u so muchh
These videos are really awesome and their quality and pedagogy are amazing. Would it be possible to have detailed video about how CRISPR Cas9 works and how it is used in genetic engineering?
Yep, videos on CRISPR are in the works - to be published soon :)
Do you plan to describe how is the whole editing system assembled for eukaryotic cells including vectors?
Yes, I do plan to talk about vectors and editing system at a conceptual level... and maybe a short "How to design CRISPR vectors" video which I am still debating.
@@theCrux oh yes, this would be a great topic to tackle!
@@theCrux Thanks it would be great! Looking forward to see it.
Thank you 😊 very much sir
Protein Expression Vectors [Part 1]: ruclips.net/video/3_WnvVgIkYo/видео.html
can you please post a video on DNA topology and supercoiling
TY!
Incredible explaination. Smart technology!
Wow u also collected information on positions of the genes Hats off Sir🥰😇
The level of conceptual clarity you have is commendable. Keep it up.❤
6:09 the [ i ] also stands for inhibitor, as it inhibits the formation of the lacZYA gene
Damn! you ARE underrated
great content!
Thank you so much ❤
You need to post more vids bro, for all the helpless students over the world
Thanks for such an informative video! Your handwriting are so clear and well-organized, making me easily understand !
Finally... Everything makes sense!!! Thank you so much! You explained all the passages very thoroughly!
Sir, Am confused about the inner and outer strand .In many books, the inner strand is heavy(H) and outer (L).So could you please exaplain?
The way you draw inner and outer does not matter - there is no such thing as inner and outer as it means nothing. H and L carry meaning because it represents the base information. As long as you follow H and L, you are good! You can flip H and L to inner and outer (and vice verse) - just make sure to keep the direction of promoters consistent.
🎉🤓🙌🏾
The eIF4 complex is recruited near the E site of the 43S(as you drew), not the A site(as you spoke 10:20)
a life save fr!!!
Great job as usual! Can you start a playlist on DNA repair system please? Or at least tell us, if possible, about your plan for the future videos. Thanks 😊
Hello! I do plan to have some videos on DNA mutation and repair in the future. At the moment, the focus is on Genetic Engineering, which I anticipate will be done by the end of this year.
Thank you so much! I struggled with this topic so much throughout my biochemistry course. You explained everything SO WELL! ❤
Thank you so much for making this!! I struggled with this mechanism and these questions so much throughout biochemistry. You explained it all SO WELL! I finally understand this replication process fully :)
Your videos are sooo helpful. Thank you so so sooooo much. It's all i need.❤
Excellent explanation.
Thank you so much, the Crux🙏 I have a week till my molecular biology finals, and now I have sets of lectures to be used as my repetitive podcast for a week. Your videos are so clear and concise, literally life saving❤ Why did I get to know about your channel just recently😢
Great job as always!
UNUSEFUL VID
Thank you so much for this video, it was very helpful!! Your explanations are clear and very well structured, what makes them very easy to follow. I also love your handwriting and drawings, they look very good! Thanks for helping me prepare for my Genetics Engeneering midterm!! :)
You are amazing!!!
Important question: why are you writing it as if the 3’ end is created first, when we both know there is ssdna looping and the 5’ end is created first because the enzyme is working in the 5’ to 3’ direction
This, with 3d videos to summarise steps in your head overall, is the best way to study genetics
I finally understood something… the primase associated with our helicase is the one making those from the lagging strand, but the leading strand only needs that one primer from the other helicase that is going to the other side, then we slide accordingly, and that’s how the other side of the primers is completed, it’s the same as the leading strand but for the other helicase
So it’s the fact that the primer creates 3’ to 5’ primers that make us need to have the polymerase act that exact way
Wait you’re saying 5’3’ but isn’t the helicase supposed to be in the side of the fork where there is a 3’ so it opens in that direction, in that case the upper helicase would be on the right and the lower one on the left. The polymerase of the lagging strand would just move according to the movement of the helicase with a lagging strand that is coiled so that when the fork progresses, the polymerase progresses away when trying to follow the complex, like shown in the famous 3d animation about dna replication, or i’m i misunderstanding the way you’re drawing it ruclips.net/video/bee6PWUgPo8/видео.htmlsi=SK0PA1cz1429eSDm
There are a couple of things to keep in context: 1) The helicase movement looks different for eukaryotes and prokaryotes. 2) The helicases cross eachother at the initiation point (usually going in opposing directions). It may become clear if you sequentially continue with the videos in my playlist on DNA replication.
@@theCrux i suppose that drawings like this are only a simplification of the actual 3d structure, and the strands are already open before the action of the helicases, which is why you didn’t draw them at their corresponding end where they are opening the dna strands, since this is a phase before that?
Amazingly Explained but I have to watch 3/4 times to grasp all the ideas, It's actually deep, Would you please make a video on bacterial engineering using arabinose operon?
this was really helpful , with all the information and notes. Thank you !!
i am a marine biologist and this video help me out to understand this theme a lot, im very glad that u make this video, ty.
Sir kya aap profesor h or aap brussels?
Very well explained , I literally searched in so many books but didn’t find proper explanation. Can you tell me your reference book though ?😊
If I manage to pass my exam Wednesday, it would probably be thanks to you
Tell me , in which book these content are given?
This was so helpful. Thank you! Preparing to transfect drosophila for the first time.
If you haven't already done it, do you think you can do a video like this about the M phase of the cell cycle, please?
There is enough word to thank you .But thank you so much sir. ❤️
I am studying for my final exams and this is finally one of the greatest video I have ever seen as a educational material. Thank you!
Arey Bhai shab gajab h gajab