I am trying to figure out how far the promoter/regulatory region of the immunoglobulin variable regions stretches upstream of the startcodon. I am aware that the conserved octamer sequence ATGCAAAT or its inverse should be present.
What is the minimal region that should be included to obtain normal expression levels, comparable to normal expression levels in B cells?
Hi Jon, someone may have more recent experience and references, but from when I worked in this area there were three elements that were highly conserved. The octamer you mentioned, and a pyrimidines rich heptamer (which could be up to 50 bases upstream of the ocatamer). in 1987 Eaton and Calame did mutational analysis to show that multiple regions were required. The paper is available for free online at
http://www.pnas.org/content/84/21/7634.full.pdf
An NCBI pubmed search will revel others who have cited this paper which may lead some more recent work in this area.
Hope this was helpful,
Frank
Thanks for the input guys! I just wanted to make sure I don't need kilobases of the upstream region to obtain something that resembles normal Ig expression. I think I will go with around 500bp and see what happens.
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