Cloning vector

This DNA and gene cloning lecture explains the properties of cloning vectors including the cloning vector uses in molecular cloning in animal and plants.
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A cloning vector is a small piece of DNA, taken from a virus, a plasmid, or the cell of a higher organism, that can be stably maintained in an organism, and into which a foreign DNA fragment can be inserted for cloning purposes.[1] The vector therefore contains features that allow for the convenient insertion or removal of DNA fragment in or out of the vector, for example by treating the vector and the foreign DNA with a restriction enzyme that creates the same overhang, then ligating the fragments together. After a DNA fragment has been cloned into a cloning vector, it may be further subcloned into another vector designed for more specific use.
There are many types of cloning vectors, but the most commonly used ones are genetically engineered plasmids. Cloning is generally first performed using Escherichia coli, and cloning vectors in E. coli include plasmids, bacteriophages (such as phage λ), cosmids, and bacterial artificial chromosomes (BACs). Some DNA however cannot be stably maintained in E. coli, for example very large DNA fragment, and other organisms such as yeast may be used. Cloning vectors in yeast include yeast artificial chromosomes (YACs).
Plasmid is an autonomously replicating circular extra-chromosomal DNA. They are the standard cloning vectors and the most commonly used. Most general plasmids may be used to clone DNA insert of up to 15 kb in size. One of the earliest commonly used cloning vectors is the pBR322 plasmid. Other cloning vectors include the pUC series of plasmids, and a large number of different cloning plasmid vectors are available. Many plasmids have high copy number, for example pUC19 which has a copy number of 500-700 copies per cell,[13] and high copy number is useful as it produces greater yield of recombinant plasmid for subsequent manipulation. However low-copy-number plasmids may be preferably used in certain circumstances, for example, when the protein from the cloned gene is toxic to the cells.[14]
Some plasmids contain an M13 bacteriophage origin of replication and may be used to generate single-stranded DNA. These are called phagemid, and examples are the pBluescript series of cloning vectors. Source of the article published in description is Wikipedia. I am sharing their material. Copyright by original content developers of Wikipedia.
Link- en.wikipedia.org/wiki/Main_Page

[@humikhan4729]

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+Humi Khan thank you. Glad you liked my lectures

[@sivakumarmalka]

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[@mayukhroy5393]

Thank You Sir, this video was very helpful.

[@pahulsingh1672]

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[@abdullahalhariri1379]

thank you for this video
do you have questions regard to this material to practice before the exam?

[@kalyanlohar476]

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[@shomusbiologyofficial]

You're welcome

[@promisendlovu8550]

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[@piko9311]

Thank you for this informative and clear tutorial!

[@vandanayadav2392]

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[@shomusbiologyofficial]

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[@kirttijapalai9622]

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[@pampaniyaarashi6641]

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[@khushisharma-pj9wu]

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[@songthanh896]

Really helpful, thanks

[@michelangelocaravaggio5018]

Hi! At 34:40 you say that the gene of interest is inserted at digestion section, between telomere sequences in circular form. this is not correct. it is digested from there to be linearized but the target gene is inserted at the part that is shown in the figure.

[@sujeongmoon7761]

plasmid has a methylase, but how they can be cut it by EcoR1..?

[@Perseus027]

Thank you very much. I solved many doubts with your video

[@shomusbiologyofficial]

Glad it helped

[@HariPrasad-ox5ri]

will the vector alter the host?

[@sonuuuu7763]

Good. No lag

[@KirtiSingh-ll5bm]

lacZ gene in plasmid....can u explain?

[@damlauzun4881]

thank you :)

[@arzoorehman1920]

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[@shomusbiologyofficial]

You're welcome

[@yonaswondimu207]

nice presentation.

[@shomusbiologyofficial]

Thank you

[@udaysinha746]

sir, the vector contains it's own DNA sequence too, besides the gene of interest .. so how is the expression of the undesired genes present originally in the vector controlled?

[@mampidutta327]

Upls answer this suman

[@TheVizkan]

the gene of interest is supplied with a strong promoter which may be activated by an inducer (eg, lactose in case of lac operon), so once the inducer is provided, the gene of interest is expressed, all other genes are not expressed

[@pankajpulate8961]

thanks to sir

[@shomusbiologyofficial]

+Pankaj Pulate you're welcome

[@JonesJinx]

If my insert is 1092 and l use the pET-28a+ plasmid vector which is 5369 bp long, how long will my recombinant vector be? Assuming l for an example use the EcoR1 RE?
thx :)

[@hasinanabi67]

thnks sir

[@misbahhussain1021]

U r great sir
Long live

[@shomusbiologyofficial]

Thank you

[@omkarkathe6598]

If we use cloning vectors for only for cloning of a gene..... Then how Tetracycline resistant gene get expressed in host cell in case of PBR322?

[@omkarkathe6598]

Is it possible to get the proteins by using cloning vectors?

[@Shuborna112]

I think in the presence of the antibiotics. The gene may become active by some activator or inducer. I'm not sure though.

[@lindaxulu7396]

Thank you :-)

[@meenakshiaggarwal6580]

Tks a lot

[@prasantapaul3424]

suman da plz ar porer videote gene expression vector r cloning vectorer differencesta banow

[@shomusbiologyofficial]

+Prasanta Paul already there

[@nimrahaseeb3440]

why it is necessary to convert mRNA to cDNA for cloning into vector

[@CrazyGamer-xi8rf]

Nimra Haseeb because we need to make copies of it... Simply inserting the mRNA would be useless

[@mouhamedosamaanbarji1896]

because the dna conatian exon and intron unlikly the mrna just have exon that's why we use the mRNA ton convert it tio cDNA

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[@sharmaprem6504]

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[@sharmaprem6504]

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[@AnonymousEasternPotato]

thank you :)