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Consider that: RNA polymerase (RNAP) is a large complex (~400 kDa in bacteria); inertia and drag would hinder its rotation. RNAP is attached to its RNA transcript, which becomes increasingly large as transcription proceeds thus increasing inertia and drag. Additionally, if RNAP were to begin to rotate around DNA, the transcript could begin to wrap around ...


4

I have generally seen the term "constitutive" used to refer to a "always on" expression of a gene with no anticipated regulatory interaction. Thus the term "constitutive knockout" seems a little bit oxymoronic to me, and I would tend to prefer a different term, like simply "knockout" or, if you really need to ...


6

I think @bob1's answer is good, and covers a lot of the bases. One thing that I think is missing however is the use of a nuclear preparation as an initial step- my understanding is that this can help with a lot of the issues of phenolic/chitinous contaminants in difficult organisms like plants/fungi/insects. Here is one suggested protocol. I would also ...


10

Insect samples are difficult to work with primarily because they contain a bunch of polysaccharides, like chitins, that are similar enough to nucleic acids that they can co-precipitate. Standard extraction procedures (e.g. Qiagen blood/tissue kits) do not handle this well without some modifications to get rid of these contaminants. The slimy aspect of your ...


4

One of the advantages you already stated that depending on protein of interest if it's already expressed in CHO cell line, researchers use that. Apart from that, this question is more of a general type. I will try my best to logically put down the things. You are right, it's more appropriate to study proteins using cultured neurons. But, there are many ...


1

Phenotype is defined as the observable physical properties of an organism (organism here includes things like cultured cell(s)). If you knock out the protein of interest you may observe a change in the function, behaviour etc. of the object you are studying via the methods you are using to look at that system. For example, in terms of proteins you might be ...


3

What are the prospects? For example, are there developments that imply the construction of logic elements and microcircuits directly inside the cell: DNA encodes the necessary proteins that are synthesized and assembled into logic gates and other components in the cell? Yes, there is an entire literature on this called "genetic circuit design", ...


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