KLONOWANIE DNA PDF

Blog. 18 December Prezi Awards The best presentations have arrived. 5 December Do this, not that: Keynote speech. 28 November Wady i Zalety Klonowania Idea 1. Idea 2. Rozmnażanie bezpłciowe – naturalne klonowanie. Klonowanie zachodzi przez: Klonowanie DNA. KLONOWANIE Klony DNA służą do: Co to jest klonowanie? Klonowanie – tworzenie genetycznej kopii fragmentu DNA, komórki lub organizmu.

Author: Grojinn Megami
Country: Central African Republic
Language: English (Spanish)
Genre: History
Published (Last): 21 February 2010
Pages: 373
PDF File Size: 13.87 Mb
ePub File Size: 20.31 Mb
ISBN: 514-7-96700-307-5
Downloads: 3193
Price: Free* [*Free Regsitration Required]
Uploader: Mell

A typical colony looks like a small, whitish dot the size of a pinhead. Plasmids and other DNA can be introduced into bacteria, such as the harmless E. So you would see things like this, which would be many, many, many cells of bacteria, there would be colonies of bacteria. Bacteria without a plasmid die.

Uses of DNA cloning. Thus, the protein is trapped in the column while other molecules from the bacteria flow through. Transformation is a key step in DNA cloning. Although, as we’ll see, it’s still quite fascinating. There are many other proteins and macromolecules floating around in bacteria besides the target protein e.

Selekcja i transformacja u bakterii

I don’t want to have to take the trouble of keep drawing the multiple strands. But when we talk about cloning and DNA cloning we’re talking about something a little bit simpler.

But maybe this colony is formed by an initial bacteria or a set of bacteria kklonowanie did not take up the plasmid so it won’t contain the actual gene in question.

This is the desired plasmid from the ligation. In one technique called affinity chromatographya mixture of molecules extracted from the lysed bacteria is poured through a columnor a cylinder packed with beads. Then, we give the bacteria a chemical signal that instructs them to make the target protein. A recombinant plasmid where the target gene is inserted after the promoter, pointing in the forward direction oriented so that it’s transcribed to make an mRNA that specified the desired protein.

These are just a few examples of how DNA cloning is used in biology today. Steps of DNA cloning.

And I’m not going to go into all the details of how you will get the insulin out and how you could make use of it, but needless to say, kllonowanie pretty cool that we can even get to this point. The molecules extracted from the cells are applied to a column that contains antibodies specific for the target protein.

  CFENGINE REFERENCE PDF

They’re bumping in dnx the right way to cause this reaction to happen then you’re taking those genes and you’re putting them with the plasmids that happen to have the right sequences at their ends so that they match up and then you also put in a bunch of DNA ligase.

Plasmids used in cloning contain an antibiotic resistance gene.

Klonowanie- Tworzenie genetycznych kopii by Klaudia Dula on Prezi

Cut open the plasmid and “paste” in the gene. Enzymy restrykcyjne i ligaza DNA. In a typical DNA cloning procedure, the gene or other DNA fragment of interest perhaps a gene for a medically important human protein is first inserted into a circular piece of DNA called a plasmid.

The bacteria are given a heat shock, which causes some of them to take up a plasmid. So how do you select for the bacteria that actually took up the plasmid? Because of these possibilities, it’s important to collect plasmid DNA from each colony and check to see if it matches the plasmid we were trying to build.

In other cases, though, the plasmid may simply close back up without taking in the geneor the gene may go into the plasmid backwards.

So this one is a good colony, put a checkmark there. And so you won’t know, hey when this bacteria, when it keeps replicating it might form one of these, it might form one of these colonies. They’re not even going to grow because there’s antibiotics mixed in with those nutrients.

Protein production and purification. For example, DNA cloning was used to build plasmids containing a normal version of the gene that’s nonfunctional in cystic fibrosis.

It occurs after restriction digest and ligation and transfers newly made plasmids to bacteria. This is not a useful plasmid. Thus, bacteria that took up the plasmid can be selected on nutrient plates containing the antibiotic. Or it can even express itself just like the genes of the organism that are in the chromosomes, express themselves. Bacteria without a plasmid will die, while bacteria carrying a plasmid can live and reproduce.

  EAGC COURSE BOOK ON COLPOSCOPY PDF

Przegląd: Klonowanie DNA

A restriction enzyme is a DNA-cutting klonodanie that recognizes a specific target sequence and cuts DNA into two pieces at or near that site. When we cut and paste DNA, it’s often possible for side products to form, in addition to the plasmid we intend to build. As an example, let’s see how DNA cloning can be used to synthesize a protein such as human insulin in bacteria.

Bacteria contain many proteins and macromolecules. See the article on restriction enzymes and DNA ligase for llonowanie more concrete example of how and why these different ligation products can form.

Several colonies are checked to identify one with the right plasmid e. Transformation and selection of bacteria are key steps in DNA klonoqanie. If the gene were backwards, the wrong strand of DNA would be transcribed and no protein would be made. In some cases, we need lots of DNA copies to conduct experiments or build new plasmids. Colonies with the right plasmid klonowqnie be grown to make large cultures of identical bacteria, which are used to produce plasmid or make protein.

Bacteria with a plasmid are antibiotic-resistant, and each one will form a colony. The promoter “points” towards the right, meaning that it will drive transcription of the DNA sequence that lies to the right.

Overview of DNA cloning. Let me write the labels down, into our plasmid that also contained a gene that gave antibiotic resistance to any bacteria that takes up the plasmid. The plasmid contains an antibiotic resistance gene, a promoter to drive gene expression in bacteria, and the target gene inserted during the ligation. Repeated nebulisation of non-viral CFTR gene therapy in patients with cystic fibrosis: And there’s a bunch of restriction enzymes, and I personally find it fascinating that we as a civilization have gotten to the point that we can find and identify these enzymes and we know at what points of DNA that they klinowanie cut.