Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/35986
標題: 百合SKP1和一花粉專一性cDNAs之特性分析
Characterization of the lily SKP1 and a pollen-specific cDNAs in Lilium longiflorum
作者: 張良棋
Chang, Liang-Chi
關鍵字: lily;百合;SKP1;pollen;花粉
出版社: 農業生物科技學研究所
摘要: 
中文摘要
百合成熟花粉的LLP13 cDNA選殖體含有兩個完整的open reading frame (ORF),命名為百合花粉SKP1 (suppressor of kinetochore protein 1 p of yeast)(LLP13-1) 和LLP13-2。北方墨點分析顯示百合花粉SKP1 mRNA長度約有600個核甘酸。核酸序列分析得知百合花粉SKP1 cDNA的長度為507 bp,具有一ORF可轉譯出127個氨基酸。百合SKP1 cDNA具有花粉專一性,在百合的根、莖、葉、花絲、雌蕊皆偵測不到SKP1 mRNA。RT-PCR進一步的証實SKP1 mRNA具有花粉專一性。在前減數分裂期、小孢子發育期和花粉成熟的早期亦偵測不到SKP1基因的表現。SKP1 mRNA的累積只發生在花粉成熟的晚期,隨著花粉/花藥的成長而有逐漸增加的趨勢。資料庫比對得知百合花粉SKP1蛋白質與小麥、水稻、阿拉伯芥,甚至人類和酵母菌的SKP1有相當高的類似性,氨基酸的相同度從26 %到34 %不等。核酸序列分析得知LLP13-2 cDNA長度為1789 bp,具有一ORF,可轉譯375個氨基酸。RT-PCR分析顯示LLP13-2只表現於花粉中,於花絲、雌蕊、花被、根、莖及葉皆無偵測到LLP13-2 mRNA,因此LLP13-2也是具有花粉專一性。在花藥發育時期上,LLP13-2 mRNA可於減數分裂和有絲分裂發生的時期和花粉成熟的晚期被偵測到。氨基酸序列比對,發現LLP13-2類似於阿拉伯芥的一個假設性蛋白質。將百合花粉SKP1轉譯區黏接到表現載體送到大腸桿菌中大量表現,可得一大小約14.5 kDa的蛋白質。

Abstract
A lily mature pollen cDNA clone LLP13 contains two open reading frames (ORF), designated as SKP1 (LLP13-1) and LLP13-2. Northern blotting analysis revealed that the mRNAs of lily SKP1 contains 600 bases. Sequence analysis of SKP1 cDNA revealed that SKP1 has 507 bp in length that encodes 127 amino acids. The expression of SKP1 cDNA is pollen -specific. No mRNA was detected in the root, stem, leaf, filament, and pistil. RT-PCR analysis also confirmed the pollen specific expression of SKP1 mRNAs. No SKP1 mRNA was detected at the stage of premeiosis, microspore development, and at the early stage of pollen maturation as well. SKP1 mRNAs were gradually accumulated to higher levels at the late stage of pollen maturation. Database search revealed that the lily SKP1 share sequence similarity to barley, rice, arabidopsis, human, and yeast SKP1 proteins, ranging from 26 % to 34 % identity. Sequence analysis of LLP13-2 revealed that LLP13-2 cDNA is 1789 bp containing an ORF that encodes 375 amino acids. The LLP13-2 is pollen specific and expressed at the occurrence of meiosis and mitosis during development and at the late stage of pollen maturation. Database search revealed that the LLP13-2 is similar to a hypothetical protein of Arabidopsis. The lily SKP1 protein was overexpressed in E. coli, from which a size of 14.5 kDa corresponding to SKP1 protien was obtained.
URI: http://hdl.handle.net/11455/35986
Appears in Collections:生物科技學研究所

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