[求救] 有關eGFP的vector問題
小弟正在讀篇有關siRNA相關paper
作者建構了一個vector，有關eGFP以及chimeric intron 我不甚了解。
以下為文中不解之處
The mir-30e precursor (98 bp) (=== 這裡是指pre-miRNA構築在vector內
amplified from the mouse genomic DNA was inserted into
the chimeric intron (36 bp 5' donor and 36 bp 3' acceptor
sequences) of an eGFP gene so that the pre-mir-30e could
be spliced from the eGFP transcript to produce the mir-
30e sister pair.
The remaining eGFP transcript would be
spliced, ligated and translated into eGFP (== 最後eGFP會自己ligate起來!?
小弟不解的是
什麼是chimeric intron (36 bp 5' donor and 36 bp 3' acceptor
sequences) 我推想他的功能可能和intron類似,是之後會被移除的地方，所以意味著
siRNA之後也會被移除出來(因為將siRNA放在chimeric intron內)
但是 36 bp 5' donor and 36 bp 3' acceptor sequences是什麼樣的方法可以讓
siRNA變成兩段小段的sequence!?!?
還是以上我都想錯了...
懇請知道的前輩提供想法 <(_ _)>
附上期刊網址 (Fig4 b 有此vector)
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=2034466
感恩再感恩 。
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