a******s
发帖数: 6 | 1 We know now that the binding between histone and DNA is specific and non-
sequence specific.Do you guys know how to prove that the histone-DNA binding
is specific, not non-specific? especially at the initial stage of this
study. I couldn't find any paper. Thanks a lot. | I*****y
发帖数: 6402 | 2 make yourself clear first
binding
【在 a******s 的大作中提到】
: We know now that the binding between histone and DNA is specific and non-
: sequence specific.Do you guys know how to prove that the histone-DNA binding
: is specific, not non-specific? especially at the initial stage of this
: study. I couldn't find any paper. Thanks a lot.
| A******y
发帖数: 2041 | 3 http://www.ncbi.nlm.nih.gov/pubmed/15100411
Not so sure what are you asking, but here is a paper should answer some of
your questions. | a******s
发帖数: 6 | 4 Thanks. I am trying to clarify my question.For most of transcription factors
, there is a specific binding site (consensus sequences) on DNA for them to
bind. Without them, no binding. We can use ChIP or EMSA to test it. However,
for histone, there is no specific binding site on DNA. But histone binds to
any DNA. So we call that the histone-DNA binding is specific, but non-
sequence specific.But for some proteins, they may have some kind of
association, but not direct binding with DNA. So my question is, how to
demonstrate that the binding between histone and DNA is a direct binding?
How the binding ability of histone to DNA was found and proved? Or in
another way to ask, can we say that the protein-DNA is direct binding if I
can show the DNA-protein binding in gel-shift assay?
【在 A******y 的大作中提到】
: http://www.ncbi.nlm.nih.gov/pubmed/15100411
: Not so sure what are you asking, but here is a paper should answer some of
: your questions.
| A******y
发帖数: 2041 | 5 Huh? You have x-ray crystal structure showing you that it is binding to the
DNA. The same paper also reconstituted the chromosome using DNA and
histone. In addition, you can see nucleosome with cryo-EM. In real
physical world, binding specificity is achieved through affinity, if you use
enough of DNA and enough random protein, they will bind except it has no
biological significance and just aggregation. To answer your question, even
BSA can shift a DNA gel if you used enough to aggregate the DNA. Shifting
at nano to pico molar range, I call it specific. |
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