黑果枸杞WD40编码基因LrAN11的克隆及表达分析

发布时间：2018-04-27 12:27

本文选题：黑果枸杞 + WD蛋白　；参考：《核农学报》2017年11期

【摘要】：WD40蛋白广泛存在于真核生物中,是一类高度保守的蛋白家族,具有广泛的生物化学和细胞生物学功能。为探索黑果枸杞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,将其命名为LrAN11,Gen Bank登录号:KY131959。生物信息学分析结果表明,该基因的编码区长1 029bp,编码342个氨基酸,蛋白分子量为38.3 kD,理论等电点为4.95,含有5个WD40基序,属于WD40类蛋白基因家族;经预测LrAN11定位于细胞质中,该基因编码的氨基酸序列具有较高的保守性,与马铃薯、番茄、甜椒和美花烟草具有较近的亲缘关系;定量RT-PCR检测表明,LrAN11基因在黑果枸杞各组织中均有表达,其中在青果中的相对表达量最高,在根和紫果中次之,而在黑果中表达最低,表明LrAN11可能参与黑果枸杞花青素的生物合成的调控;在韩国枸杞和0901枸杞品种中表达显著,在其他14个枸杞品种中表达均较低且无显著差异性(P0.05),说明LrAN11的表达具有品种特异性。此外,随着NaCl处理时间的延长,该基因的表达量先降低后升高,且在2、12和24 h的表达量与对照相比存在显著差异性(P0.05),说明LrAN11可能参与黑果枸杞对盐胁迫的响应。本研究结果为进一步阐明黑果枸杞LrAN11基因的功能及其在黑果枸杞遗传改良中的应用奠定了基础。
[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.
【作者单位】：宁夏大学生命科学学院;宁夏林业研究院种苗生物工程国家重点实验室;
【基金】：宁夏自然科学基金项目(NZ16215) 国际合作项目(2015DFA90900)
【分类号】：Q943.2;S567.19

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