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如何製造基因轉殖果蠅?

所謂的基因轉殖果蠅,是指某一種果蠅品系,他的genome裡面就帶有外來的基因,非果蠅自己原本就有的。像是GAL4/UAS binary expression system裡的GAL4 lines, UAS lines, LexA lines ...etc. (可以說…做實驗用的果蠅,絕大部分都是基因轉殖果蠅)
再來,怎麼將外來的基因,鑲嵌到果蠅的genome裡面,讓它可以世世代代遺傳下去呢?主要是利用轉位子transposon的神奇力量 XD(1983 Barbara McClintock 因從玉米發現轉位子
 而獲得諾貝爾獎) 某些生物體內,原本就有Transposon,像玉米、人類也有。果蠅目前最流行的,就是P element.

P element其實不是外來的東西(不像GAL4/UAS system,那是從yeast搬過來果蠅用的),只是做實驗用的果蠅,都經過清理,體內都不帶有這種transposon. Wikipedia寫得很清楚,請看http://en.wikipedia.org/wiki/P_element

然後,P element其實有分可以自己跳來跳去的autonomous和不能自己跳的nonautonomous
(nonautonomous個人覺的…就是閹割版嘛~)
不過,重點是…transposase可以辨識p element兩端的sequence(5'p and 3'p),然後將兩端sequence連帶中間的片段切離genome,然後再找地方插回去。(PS:這個過程不是完美的,有一定的機率會出ㄘㄟˋ,留下部分的p element片段,或是拉走附近genome的一整段造成突變)

##產生基因轉殖果蠅的方法,就是將transposase和一段P element的片段,同時打入embryo stage的生殖細胞附近。如此一來,這個embryo長成成蟲後,他的精子(她的卵)的genome就會有機會帶有這段外來的P element。##

但怎麼知道哪些才是帶有插入的P element呢?這個被打的embryo,是白眼突變果蠅;而外來的P element片段裡,帶有讓果蠅眼色回復正常紅色的基因。所以後來如果眼色回復成紅色,就代表P element有插入該品系的genome裡面。

Mx

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