Expression vector

This lecture explains about the expression vector properties and the uses of expression vector in gene cloning.
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An expression vector, otherwise known as an expression construct, is usually a plasmid or virus designed for protein expression in cells. The vector is used to introduce a specific gene into a target cell, and can commandeer the cell's mechanism for protein synthesis to produce the protein encoded by the gene. The plasmid is engineered to contain regulatory sequences that act as enhancer and promoter regions and lead to efficient transcription of the gene carried on the expression vector.[1] The goal of a well-designed expression vector is the production of significant amount of stable messenger RNA, and therefore proteins. Expression vectors are basic tools for biotechnology and the production of proteins, for example insulin which is important for medical treatments of diabetes.
An expression vector has features that any vector may have, such as an origin of replication, a selectable marker, and a suitable site for the insertion of a gene such as the multiple cloning site. The cloned gene may be transferred from a specialized cloning vectors to an expression vector, although it is possible to clone directly into an expression vector. The cloning process is normally performed in Escherichia coli, and vectors used for protein expression in organisms other than E.coli may have, in addition to a suitable origin of replication for its propagation in E. coli, elements that allow them to be maintained in another organism, and these vectors are called shuttle vector.
Elements for expression
Further information: Transcription (genetics) and Translation (biology)
An expression vector must have elements necessary for protein expression. These may include a strong promoter, the correct translation initiation sequence such as a ribosomal binding site and start codon, a strong termination codon, and a transcription termination sequence. There are differences in the machinery for protein synthesis between prokaryotes and eukaryotes, therefore the expression vectors must have the elements for expression that is appropriate for the chosen host. For example, prokaryotes expression vectors would have a Shine-Dalgarno sequence at its translation initiation site for the binding of ribosomes, while eukaryotes expression vectors would contain the Kozak consensus sequence.
The promoter initiates the transcription and is therefore the point of control for the expression of the cloned gene. The promoters used in expression vector are normally inducible, meaning that protein synthesis is only initiated when required by the introduction of an inducer such as IPTG. Protein expression however may also be constitutive (i.e. protein is constantly expressed) in some expression vectors. Low level of constitutive protein synthesis may occur even in expression vectors with tightly controlled promoters.
Protein tags
Main article: Protein tag
After the expression of the gene product, it is usually necessary to purify the expressed protein. However, separating the protein of interest from the great majority of proteins of the host cell can be a protracted process. To make this purification process easier, a purification tag may be added to the cloned gene. This tag could be histidine (His) tag, other marker peptides, or a fusion partners such as glutathione S-transferase or maltose-binding protein. Some of these fusion partners may also help to increase the solubility of some expressed proteins. Other fusion proteins such as green fluorescent protein may act as a reporter gene. Source of the article published in description is Wikipedia. I am sharing their material. Copyright by original content developers of Wikipedia.
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You are seriously amazing. No other videos explain the details that you touch up on. Literally helping me understand my dissertation. I don't know what I would have done without your videos. Thank you!

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[@tinyikonkuna1479]

I had no clue how the expression was connected to the central dogma, thank you so much, sir . May God richly bless you.

[@shomusbiologyofficial]

Thank you

[@paridhi796]

Jo concept university prof ni achhe se samjhate wo concept aap bahut hi fundamental tareeke se samjhadete ho sir

[@shomusbiologyofficial]

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[@anamikarajput1347]

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[@Alan-ul5qw]

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[@shomusbiologyofficial]

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[@arzoorehman1920]

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[@shomusbiologyofficial]

Glad to hear that you're getting benefit from my lectures

[@deyvanaiarumugam6083]

Thank you for your great explanation. You made me to understand precisely.

[@b2bb461]

Thank you so much my favorite biology teacher. Would you please make several videos about snap gene soft ware? It is not only beneficial, but also related to cloning lectures.

[@shomusbiologyofficial]

Okay I will try

[@b2bb461]

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[@kianagoudarzi1619]

Thank you for putting so much time into your tutorials. Great details. Extremely helpful🙏🏻

[@meli260582]

pBR322 is a cloning vector and you put it to the expression vector column, whereas pBAD/D- Topo is an expression vector and you placed it to the cloning vector column. It would have been better if you just reduced the font of the expression vector features and put the right vector there. Just by simply looking at this slide is misleading.

[@DAK1985]

your lectures are really nice. Can you please lecture about plasmid mapping and the direction of the arrows

[@Cacarot488]

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[@TheJonas1014]

Jasna

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Sir This video was very Helpful.

[@shomusbiologyofficial]

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[@shomusbiologyofficial]

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[@kalyanlohar476]

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[@shomusbiologyofficial]

You're welcome

[@wonderfulIndia1]

Sir expression vectors k kon s examples h . Pbr322 . Puc10 . Bacteriophage . Cosmid phasmid. Bac yac ye sb cloning vector k example h ? Plzz reply don't ignore 🙏

[@nirjapatel1540]

I love your videos. They are very informative. Can you please make a video on Woodchuck Hepatitis Virus (WHP) Posttranscriptional Regulatory Element (WPRE) genes, with its basic concepts, usage and functionality? Thank you!

[@shomusbiologyofficial]

Glad to hear that you're getting benefit from my lectures

[@4Getf00l]

What does "copy number of gene" mean? and, for a specific gene, is it possible to check its copy number online, for example from NCBI?

[@user-ry1cc1im6f]

When we use E.coli strains for protein expression we use Rosetta which has chloramphenicol resistance Gene which blocks protein production. But since the E.coli is already resistant why we add also chloramphenicol?

[@juliaandreunoguera9022]

Very helpful, thanks!

[@shomusbiologyofficial]

You're welcome

[@jatrokulani3371]

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[@Bodwaypatt]

You could do a video of Promoters used in protein expression (lac, trp, tac, PL, Pt7...) in yeast, Pichia pastori, and baculovirus

[@marwanmohamed6575]

can some one explain how the tags like histadine tags is going to be expressed,, isnt our gene already got a stop codon?

[@sasidharankr2344]

The tag sequence can be present before stop codon....

[@vijayabharathigopi261]

can you refer any books for this expression vectors to study in detail

[@nasreenfatma6735]

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[@priyankah.t523]

Thank you so much

[@shomusbiologyofficial]

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[@yoginiilame8718]

Thank you

[@tayebwadickson5165]

thanks alot

[@himaniparmar8377]

Thank you sir

[@ISSOC1]

Great!!!

[@shomusbiologyofficial]

You're welcome

[@meli260582]

And don't waste too much time changing the stupid colors!

[@pankajpulate8961]

thank you so much sir

[@shomusbiologyofficial]

+Pankaj Pulate glad to hear that