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If alleles occur at the same locus on a chromosome, does that mean they are of the same size? I was under the impression that they were, but I saw a gel electrophoresis testing different alleles of the same gene and saw them separate. I don’t understand how they could separate if they are of the same size, but if they aren’t, then how can they be housed in the exact same locus?

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  • $\begingroup$ Are you sure it is just a normal gel electrophoresis? It could be some technique like RFLP which can differentiate alleles. $\endgroup$ – WYSIWYG Mar 26 '16 at 14:48
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    $\begingroup$ If they are not on the same locus, then you can find them using electrophoresis. Basically you are checking for copy number. However, alleles at same locus can be of different sizes (but I don't think the differences are that great, in most cases). There are a lot of possibilities but unless you provide details, it is hard to explain what exactly you observed in your experiment. $\endgroup$ – WYSIWYG Mar 26 '16 at 14:53
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In general, alleles don't have to be the same size. Two major examples which come to mind are the Huntingtin gene and FMR1.

Huntingtin is the causative gene of Huntington's disease. In people with Huntington's, a sequence of three nucleotides is repeated. The number of repeats varies, from a low of 9 in unaffected individuals, to more than 60 in severely affected ones. So depending on how many repeats you have, each of your Huntingtin alleles may have a different nucleotide size.

FMR1 is the causitive agent of Fragile X syndrome. Like Huntingtons, the gene varies in the number of trinucleotide repeats, but for FMR1, affected individuals can have over 1000 repeasts, meaning an extra 3 kilobases of genetic material versus the alleles of an unaffected individual.

Other genetic disorders are also related to the difference in allele size. You can have a deletion of a region of DNA (or an insertion) which is normally silent (recessive). The normal-length allele compensates for the defective allele, but the differently-sized disease allele is still present in the population, and counts as an allele of the gene.

It's not just disease-prone alleles which exhibit length polymorphism. A number of other genes will show variation on both the DNA and on the protein level. If the insertion/deletion is a multiple of three nucleotides and occurs in a loop region of the protein or on an intrinsically disordered domain, the insertion has a reasonable chance of minimal structural effect on the expressed protein. For one (completely arbitrary) example, PER3 has two major length variants in the population, with a 54 base pair difference between the two. Different populations have vastly different prevalence for each, with a range of 20-90% for the shorter version.

One thing to keep in mind is that "locus" points are just a human convention to help us map DNA. To the cell, DNA is just a single long molecule. (Or multiple molecules, for different chromosomes.) There isn't any labels for absolute position in the cell, so inserting or deleting a chunk of DNA mainly affects just the local environment. There isn't any sort of long-range indexing that gets messed up by the insertion.

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  • $\begingroup$ So what exactly happens when the repeats increase? Does this gene code for a specific enzyme? Do the increased repeats change the enzyme activity? $\endgroup$ – curious_cat Mar 27 '16 at 3:44
  • $\begingroup$ @curious_cat the exact function of huntingtin is not really known. In most of these poly-Q proteins, the repeat is not present in any functional (binding/catalytic) regions. This ideally should not affect their activity but there are other factors at play here. The exact mechanisms of poly-Q toxicity are still not clear: one thing that is known is that this causes a stress response. $\endgroup$ – WYSIWYG Mar 27 '16 at 5:48
  • $\begingroup$ @WYSIWYG What about in general? Say alleles that determine other phenotypic features, say eye color or sickle cell anemia or hemophilia etc. How do the difference in amino acid repeats / type lead to the feature difference. What is the mechanism. Is it a specific enzyme's activity alone or something else? $\endgroup$ – curious_cat Mar 27 '16 at 5:57
  • $\begingroup$ @curious_cat usually the differences in alleles (at least the coding region of mRNA) are mostly substitutions (which can lead to a change in the activity). Indels are dangerous as they can disrupt the reading frame. Amino acid repeats like in poly-Q diseases arise because of homopolymeric expansion of the CAG triplet. As I said, how poly-Q affects the activity of the affected protein is not fully known. Moreover, different polyQ diseases have diverse kinds of affected proteins ranging from androgen receptor to TATA-binding protein. $\endgroup$ – WYSIWYG Mar 27 '16 at 6:17
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    $\begingroup$ @OneFace The disease causing variants are the most obvious, as they're the most studied and have substantial length variation. But even in asymptomatic individuals there's a variation in length for Huntingtin and FMR1. Variations in other, non-disease-associated genes exist too, and I added an example I found from a quick literature search. $\endgroup$ – R.M. Mar 28 '16 at 19:29
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One possibility of what you were looking at was a nuclease digestion experiment. A subject's gene sequence could be amplified from their DNA using PCR, giving a band of a certain size when electrophoresed on a gel. The PCR'd product could then be digested with another nuclease that specifically cuts one allelic variant, but not another. For example, check out this rather unrefined image I made:

fake gel

Lane 1 is the molecular weight ladder. Lane 2 shows the PCR product of our gene of interest, which has two possible alleles - one that can be cut with our nuclease, and one that can't. The rest of the lanes show the result of incubating the PCR product from different individuals with our nuclease. Lane 3 shows a homozygote for one allele, the one that cannot be cut. Lane 4 shows a heterozygote, where the uncut 900 bp band is also seen with the cut 400 and 500 bp fragments. Lane 5 shows a homozygote carrying the cuttable allele on both chromosomes - there is no intact 900 bp band.

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  • $\begingroup$ This does provide an example of why alleles could run differently on a gel, but fails to answer the underlying question about allele size $\endgroup$ – Luigi Mar 26 '16 at 19:04

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