[DIYbio] Re: Plasmids as a vector

2:11 PM |

A lot of times vector and plasmid are used as interchangeable terms (although they do refer to different things) because almost all vectors are plasmids. There are a few different vectors, however, like viral vectors. Even those are usually based on plasmids. Almost all plasmids are vectors. A vector usually refers to something that transmits something from one organism to another, like a vector for disease. A plasmid on the other hand is a piece of DNA that replicates independently inside of a cell. Scientists use plasmids to transmit cloned information from one organism to another, so that's why they're almost always called vectors. (Just how I think of the terminology)

Most scientists use E coli, which the plasmid for cloning is the 'vector' for gene transformation. Since you can just use a straight up expression plasmid as an expression vector, nothing really needs to be done. However some organisms don't have plasmids, and that's where you want to use integration. (talking expression vectors here)


Integration of parts into the genome is sometimes more difficult, such as integration into E coli. Lambda red (in plasmid pKD46) works well for that organism. That's probably what the biobrick people were using. Other organisms, like S cerevisiae, are even better at integration and will readily integrate any DNA you have (given homologous sites). pGREEN is a plant vector I know of that I think integrates into the genome, but I'm not sure. Might want to investigate further on that one.

ORF stands for "open reading frame", also known as a protein. Most often you want to add in your own ORF or protein.

-Koeng



On Saturday, February 7, 2015 at 2:08:55 PM UTC-8, Louis Weber wrote:

Hey Guys

First of all sorry for my bad English. I`m a beginner and need some help about plasmid.

So you can use plasmids for cloning (to copy a DNA-Part many time in a Bacteria), but also as a vector for gene transfection or transformation is that right?

So for Example, can I design a Vector and put it with for example with electroporation in the cell, and the Cell would, if I had de right Promoter, transcript the DNA and produce the Protein that I had on the Vector?

If that's right, I had two questions…

1 How can I bring the Vector that is in the cell plasma because of de Electroporation into the Genome of the Cell?

2 I have seen the website http://www.addgene.org/ there it gives many plasmid. Are this also vectors? And in there Plasmid it has Promoters, Parts for restriction enzyme and ORFs. But is in the ORFs already some codes for Proteins or must I add this? And can I find there a vector for plants?

Sorry for my  questions but I am a beginner and found not very much information on the internet.

Louis

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