红果醋栗RrFAD2基因的克隆与表达分析

Cloning and Expression Analysis of Rr FAD2 Gene in Ribes rubrum

本研究克隆到红果醋栗(Ribes rubrum L.)脂肪酸脱氢酶2(co-3 fatty acid desaturase,FAD2)基因cDNA全长序列,命名为RrFAD2,GenBank登录号为MT795718。RrFAD2全长1470 bp,开放阅读框序列全长1152bp,编码383个氨基酸。生物信息学分析表明,RrFAD2蛋白具有6个跨膜结构域,分子量为43.92 kD,等电点为8.93。利用荧光定量PCR分析RrFAD2基因在红果醋栗不同组织和种子发育中的表达。结果表明该基因在根、茎、叶、花、果实、种子中都表达,在种子中表达量最高,花和根中最低,具有组织特异性。在种子发育过程中,RrFAD2基因的表达先上升后下降,而种子成熟的晚期(花后80 d左右)表达量达到最高。该研究为解析红果醋栗RrFAD2基因功能和研究醋栗不饱和脂肪酸合成提供理论参考。

The study cloned the co-3 fatty acid desaturase 2(FAD2)gene from Ribes rubrum L.,which was designated as RrFAD2 with accession number of MT795718 in the NCBI GenBank.The full cDNA sequence of the RrFAD2 gene has 1470 base pairs,containing an open reading frame of 1152 bp encoding a protein of 383 amino acid.Bioinformatics analysis showed that RrFAD2 protein has 6 transmembrane domains with a molecular weight of 43.92 k D and an isoelectric point of 8.93.We determined the expression of the RrFAD2 gene in different tissues and the process of seed development by quantitative-PCR analysis.The results showed that the gene was expressed in root,stem,leaf,flower,fruit and seed,with the highest expression in seed and the lowest in flower and root,which has tissue specificity.In the process of seed development,the expression of RrFAD2 gene in creased first and then decreased,and the expression was highest in the late stage of seed maturity(about 80 days after florescence).These results provided a theoretical reference for analyzing the function of RrFAD2 gene in red currant and the synthesis of unsaturated fatty acids in currant.