水稻OsDEADl-1基因的功能性探討

Functional analysis of OsDEADl-1 gene in rice

黃俊凱(Chun-Kai Huang)

指導教授 ： 陸重安

國立中央大學/理學院/生命科學系/碩士(2006年)

摘要

在此研究中，我們得到了一株水稻的轉殖品系 Osdeadl-1 (Osdead box protein like-1) 的轉殖株，而此轉殖株並沒有同型合子的突變株後代。 OsDEADl-1比對分析結果是屬於DEAD box RNA helicase的一員，在生理功能上RNA helicase解開雙股RNA，確保RNA形成正確的型式以執行其正確的功能。 OsDEADl-1 cDNA包含1506去氧核醣核酸。藉由RT-PCR分析中OsDEADl-1在不同水稻組織中皆有表現。藉由綠螢光蛋白 (GFP) 標定在OsDEADl-1上，在洋蔥表皮細胞及水稻癒傷組織的原生質體中清楚觀測GFPOsDEADl-1只有在細胞核中表現。同時我們在阿拉伯芥中找到AtDEADl-1， AtDEADl-1與OsDEADl-1有77%相似度，在 Atdeadl-1突變株的T3種子亦無法得到同型合子的突變株，顯示AtDEADl-1及OsDEADl-1突變後將造成阿拉伯芥及水稻致死的外表型。並藉由RT-PCR分析結果AtDEADl-1在各個不同的組織中也皆有表現。在Osdeadl-1及Atdeadl-1突變株子代的基因型分析時，發現野生型植株比上異型合子的植株比例為1:1，最後於AtDEADl-1 異型合子突變株的配子與野生型阿拉伯芥 (Len.)正反交的實驗中，顯示Atdeadl-1異型合子突變株的雄配子體可能在其受精或雄配子發育上有缺陷，所以造成無法產生同型合子的突變株後代。

關鍵字

轉殖 ； 水稻 ； 解旋酶

並列摘要

In this study, we had a mutant line Osdeadl-1 (Osdead box protein like-1). The Osdeadl-1 mutant didn’t find any homozygous mutant progeny. In blast analysis, OsDEADl-1 is a member of the DEAD box RNA helicase. RNA helicase has biological function to unwind double strand RNA to assure the correct foldings of RNA molecules to execute the correct biological functions. OsDEADl-1 cDNA includes 1506 nucleotides. The OsDEADl-1 expresses in all tissues in Oryza sativa by RT-PCR analysis. By green fluorescent proteins fused with OsDEADl-1, we clear observe GFPOsDEADl-1 expressed in the nucleus in the onion epidermal cell and rice callus protoplast. At the same time, we found AtDEADl-1 in Arabidopsis thaliana. AtDEADl-1 and OsDEADl-1 have 77% similarity. In the population of the T3 seeds of Atdaedl-1 also can’t find homozygous mutant. The data reveals the AtDEADl-1 and OsDEADl-1 mutant is lethal phenotype in Arabidopsis thaliana and Oryza sativa. And by RT-PCR result, AtDEADl-1 gene expresses in different tissue in Arabidopsis thaliana. In genotype analysis of the progeny of the Osdeadl-1 and Atdeadl-1 were obtained a ratio of 1:1, as heterozygote to the wild type. Last, in the gametophytes of Atdeadl-1 heterozygous mutant to cross with the wild type (Len.) suggests male gametophyte may have defect in its gametophyte development or its insemination. This is the reason we can’t get homozygous mutant.

並列關鍵字

transformation ； helicase ； rice

參考文獻

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