本文選題：黑果枸杞 + WD蛋白��； 參考：《核農(nóng)學(xué)報(bào)》2017年11期

【摘要】：WD40蛋白廣泛存在于真核生物中,是一類高度保守的蛋白家族,具有廣泛的生物化學(xué)和細(xì)胞生物學(xué)功能。為探索黑果枸杞LrAN11的功能,采用同源克隆的方法克隆了黑果枸杞WD40蛋白基因,將其命名為LrAN11,Gen Bank登錄號:KY131959。生物信息學(xué)分析結(jié)果表明,該基因的編碼區(qū)長1 029bp,編碼342個(gè)氨基酸,蛋白分子量為38.3 kD,理論等電點(diǎn)為4.95,含有5個(gè)WD40基序,屬于WD40類蛋白基因家族;經(jīng)預(yù)測LrAN11定位于細(xì)胞質(zhì)中,該基因編碼的氨基酸序列具有較高的保守性,與馬鈴薯、番茄、甜椒和美花煙草具有較近的親緣關(guān)系;定量RT-PCR檢測表明,LrAN11基因在黑果枸杞各組織中均有表達(dá),其中在青果中的相對表達(dá)量最高,在根和紫果中次之,而在黑果中表達(dá)最低,表明LrAN11可能參與黑果枸杞花青素的生物合成的調(diào)控;在韓國枸杞和0901枸杞品種中表達(dá)顯著,在其他14個(gè)枸杞品種中表達(dá)均較低且無顯著差異性(P0.05),說明LrAN11的表達(dá)具有品種特異性。此外,隨著NaCl處理時(shí)間的延長,該基因的表達(dá)量先降低后升高,且在2、12和24 h的表達(dá)量與對照相比存在顯著差異性(P0.05),說明LrAN11可能參與黑果枸杞對鹽脅迫的響應(yīng)。本研究結(jié)果為進(jìn)一步闡明黑果枸杞LrAN11基因的功能及其在黑果枸杞遺傳改良中的應(yīng)用奠定了基礎(chǔ)。
[Abstract]:WD40 protein is a kind of highly conserved protein family, which has a wide range of biochemistry and cellular biological functions. In order to explore the function of LrAN11, the WD40 gene of Lycium barbarum was cloned by homologous cloning and named LrAN11G Bank accession number: KY131959. The results of bioinformatics analysis showed that the coding region of the gene was 1029 BP, encoding 342 amino acids, the molecular weight of the protein was 38.3 KD, and the theoretical isoelectric point was 4.95, which contained five WD40 motifs and belonged to the family of WD40 protein genes, and LrAN11 was predicted to be located in the cytoplasm. The amino acid sequence encoded by this gene was highly conserved and had close relationship with potato, tomato, sweet pepper and tobacco, and the quantitative RT-PCR analysis showed that LrAN11 gene was expressed in all tissues of black fruit Lycium barbarum L. The relative expression was the highest in the green fruit, followed by the root and purple fruit, and the lowest in the black fruit, indicating that LrAN11 may be involved in the regulation of the biosynthesis of anthocyanin in the black fruit medlar, and in the Korean Lycium barbarum and 0901 Lycium barbarum varieties, the expression was significant. The expression of LrAN11 in 14 other Chinese wolfberry cultivars was low and had no significant difference (P0.05A), which indicated that the expression of LrAN11 was varietal specific. In addition, with the prolongation of NaCl treatment time, the expression of the gene decreased first and then increased, and there was a significant difference between the two groups at 2h and 24h, indicating that LrAN11 might be involved in the response of Lycium barbarum L. to salt stress. The results of this study laid a foundation for further elucidating the function of LrAN11 gene and its application in genetic improvement of black medlar.
【作者單位】： 寧夏大學(xué)生命科學(xué)學(xué)院;寧夏林業(yè)研究院種苗生物工程國家重點(diǎn)實(shí)驗(yàn)室;
【基金】：寧夏自然科學(xué)基金項(xiàng)目(NZ16215) 國際合作項(xiàng)目(2015DFA90900)
【分類號】：Q943.2;S567.19

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