探討mir-100對於果蠅蛹期存活率的影響

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詹家瑋(Chia-Wei Chan) 查詢紙本館藏

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論文名稱

探討mir-100對於果蠅蛹期存活率的影響
(mir-100 does not influence pupal survival rate in Drosophila melanogaster)

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至系統瀏覽論文 (2025-5-31以後開放)

摘要(中)

在果蠅（D. melanogaster）中，已知mir-100 會在果蠅發育時的前蛹期
開始表現，先前研究發現在Canton S 品系中的mir-100 表現量較其他品系果
蠅來的低，且蛹期存活率遠低於其他品系的果蠅，而且該品系基因組中mir-
100 下游有一段flea 跳躍因子的存在，推測其可能影響了mir-100 的表現
量。本論文研究延續前人研究結果，測試了mir-100 primary transcript 的轉
錄，在RT-PCR 的實驗結果中發現帶有flea 跳躍因子的果蠅，其mir-100 所
在的let-7 cluster 在primary transcript 的過程中會被分成兩個或以上的片段。
接著利用遺傳背景控制與miRNA sponge 的方法檢測mir-100 對蛹期存活率
的影響。在置換過染色體且具有flea 跳躍因子的果蠅，其蛹期存活率顯著地
低於其他品系，但是利用抑制mir-100 表現量之品系的果蠅測試其蛹期存活
率，結果發現其蛹期存活率並無明顯差異。此兩種方法所測得mir-100 對於
果蠅蛹期存活率的影響不同，需更進一步的實驗來驗證。

摘要(英)

In Drosophila melanogaster, it is known that the expression of mir-100 starts at the
pre-pupal stage. The expression level of mir-100 in a strain isolated from Canton S
strain, named CS-UCI, is lower than that of other strains and the survival rate of
this strain at the pupal stage is much lower than that of other strains, as found in
the previous studies. The previous study revealed the presence of a flea transposon
insertion in the downstream of mir-100, presumably affecting the expression level
of mir-100. In this study, the primary transcript of mir-100 was examined by RTPCR.
Based on the results of three RT-PCR experiments, the let-7 cluster,
including mir-100, was transcribed into more than two primary transcripts in CSUCI
flies. Next, the chromosome replacement of X and 3rd chromosomes in CSUCI
and control strains was carried out to control the genetic background and the
pupal survival rate was tested at 25°C . The results showed that flies carrying flea
insertion exhibited lower survival rate at the pupal stage in comparison of other
strains. However, the pupal survival rate did not change when the miR-100
expression level was decreased by mir-100 sponge. Further experiments on
measuring mir-100 expression level in genetic background controlled flies and
mir-100 sponge flies are needed to resolve the contradictory results in pupal
survival rate.

關鍵字(中)

★ 果蠅
★ 轉錄
★ mir-100
★ flea 跳躍因子

關鍵字(英)

★ Drosophila melanogaster
★ primary transcript
★ mir-100
★ flea transposon

論文目次

目錄
中文摘要 ………………………………………………………………… i
英文摘要 ………………………………………………………………… ii
誌謝 ………………………………………………………………… iii
目錄 ………………………………………………………………… iv
表目錄 ………………………………………………………………… vi
圖目錄 ………………………………………………………………… vii
一、緒論…………………………………………………………… 1
1-1 MicroRNA 的簡介…………………………………………… 1
1-2 MicroRNA cluster 與其基因表現的調節…………………… 2
1-3 研究miRNA 功能的方法…………………………………… 5
1-4 研究動機……………………………………………………… 6
二、研究方法……………………………………………………… 7
2-1 果蠅飼養與染色體置換流程………………………………… 7
2-2 總去氧核醣核酸(Total genomic DNA)萃取與let-7 cluster 區
段檢測…………………………………………………………
7
2-3 總核醣核酸(Total RNA)萃取與RT-PCR…………………… 9
2-4 果蠅蛹期期存活率試驗……………………………………… 10
2-5 統計分析方法………………………………………………… 11
三、結果…………………………………………………………… 12
3-1 染色體置換後的檢測………………………………………… 12
3-2 flea 跳躍因子會隨著let-7 cluster 一起轉錄出來…………… 13
3-3 果蠅蛹期存活率之比較……………………………………… 14
四、討論…………………………………………………………… 15
4-1 flea 可能影響果蠅之蛹期存活率及mir-100 的表現量……… 15
4-2 flea 隨著let-7 cluster 轉錄可能造成的影響………………… 15
4-3 檢討CS-UCI6 與mir-100 sponge 兩組蛹期存活率的實驗為
什麼結果不同…………………………………………………
16
v
五、結論…………………………………………………………… 18
六、參考文獻……………………………………………………… 19
vi
表目錄
表一研究中使用的果蠅品系……………………………………… 23
表二研究中使用的引子…………………………………………… 24
vii
圖目錄
圖一 microRNA 的生合成………………………………………… 25
圖二果蠅染色體置換流程………………………………………… 26
圖三 let-7 cluster 區段與引子示意圖……………………………… 27
圖四染色體置換後的檢測………………………………………… 28
圖五 flea 跳躍因子無法隨著let-7 Cluster 一起轉錄出…………… 29
圖六 flea 跳躍因子會隨著let-7 一起轉錄出……………………… 30
圖七 Pri-mir-100 在各品系中可以正常被轉錄出來……………… 31
圖八培養箱25℃環境下的蛹期存活率差異……………………… 32
圖九檢測染色體置換後的果蠅蛹期存活率差異………………… 33
圖十利用microRNA sponge 檢測mir-100 對於果蠅蛹期存活率
的影響…………………………………………………………
34

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指導教授

葉淑丹(Shu-Dan Yeh)

審核日期

2020-8-17

推文