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4

The answer is not simple - @shigeta mentioned a few mechanisms leading to single gene-to-multi protein relationships - and the answer is certainly not short (there are thousands of these genes). But anyway "alternative splicing" seems to be the primary mechanism according to this article, so rather than listing all alternatively splicing genes, here are the ...


4

There are a large number of ways a protein variant can be produced by post translational modification. The question may seem obvious, but its really quite broad. I can start this out. I doubt I know all the ways a single transcript can produce variant proteins. A detailed description might be more like a review article than an answer here. First, ...


3

Just for balance, here is an example of a single protein being constructed from two primary gene products (two separate genes involved) via protein splicing.


3

Not sure that I have ever come across trace metals in recipes (I assume that you aren't referring to the Ca/Mg salts component of M9), but it is certainly the case that many K12 strains (e.g. C600, DH5α) require thiamine because they are thi mutants. This doesn't apply to BL21 since it is from a different lineage (it is a B strain). In a lab where ...


2

The sequences you have posted seem to be (protein) amino acid sequences. The stop codon are present in DNA sequences and in mRNA sequences. In DNA, the bases are A, G, C and T; stop codons are TAG, TGA and TAA. In RNA, the bases are A, G, C and U; stop codons are UAG, UGA and UAA. In DNA and RNA, other letters are used to specify degeneracy. What you have ...


2

I think that enzyme-linked immunosorbent assay (or ELISA) is the best way to do so, given you have an antibody against the protein to coat the sample wells with... You could use mice to create polyclonal antibodies against your protein by injecting the protein and collecting the serum and purifying it...as long as there isn't similar proteins to the ...


1

Verify where these transcripts are generally present (this should not be difficult for common organisms) If they are all expressed in a certain tissue use cDNA from that tissue If not then pool the cDNAs from different tissues such that all genes can be amplified cDNA libraries (including whole organism) are also available and you may obtain them from the ...


1

Things can certainly vary for different ligands and different levels of cascading but these are the reactions that you may consider. Receptor activation Nuclear translocation (lets assume it is just a part of the receptor. You can assume secondary messengers; it will add an extra step) DNA-binding and Transcription activation Transcription Nuclear export ...


1

In general, leupeptin (a cysteine, threonin and serine protease) and aprotinin (sometimes called Trasylol; trypsin inhibitor) are included in most mammalian cell lysate buffers. Whether you need them is really up to your protein of interest: if you know how it gets degraded in vitro, then you should be fine with inhibition of just those proteases. If instead ...


1

The Wikipedia article on epistasis has a section on epistasis within genes, which it terms intragenic complementation: Just as mutations in two separate genes can be non-additive if those genes interact, mutations in two codons within a gene can be non-additive. In genetics this is sometimes called intragenic complementation when one deleterious mutation ...


1

There is a paper for transfection of mammalian cells here which has a bit of comparison of stop codon and protein yield. Otherwise this article suggests having a 'rare' codon after the stop to prevent readthrough. But yes, I'm not sure why one wouldn't simply put two stops after each other.


1

I used pEYFP-N1 (stock vector) where the spacing between the CMV promoter and the EYFP start codon is ~88 nt, and the expression is high. I also cloned a protein 5' of the EYFP, only 4 nt after the CMV promoter. The expression of this fusion was also high. Because I was only using the chimeric protein in experiments, I did not try to assess differences in ...


1

You can use a bidirectional promoter. The problem that you mentioned about proteins not expressed in same level happens because of competition for polymerase. But there are well optimized parts and also commercially available vectors that work fine. You can clone genes in a serial order. It won't be a problem. Just leave a 100bp linker after the polyA ...



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