摘要
为获得龙眼PAL基因的序列,并研究该基因在转录水平的表达,采用反转录PCR获得PAL的保守序列,然后利用RACE法克隆PAL基因的3'cDNA和5'cDNA序列,再利用DNAMAN软件拼接获得PAL基因的cDNA全长序列。以actin基因作为内参进行实时荧光定量PCR,研究PAL基因在4个龙眼品种嫩叶中的表达差异。结果表明,克隆得到Dlpal基因的cDNA全长序列为2 532 bp,开放阅读框为2 101 bp,编码839个氨基酸,分子量为92.259 ku,等电点为7.38。BLAST比对结果显示,该序列与其它物种的PAL基因具有较高的同源性。实时荧光定量PCR结果表明,Dlpal基因在4个龙眼品种‘乌龙岭’、‘松风本’、‘立冬本’和‘石峡’的嫩叶间的表达量差异显著,其中在‘立冬本’中表达量最高,在松风本中表达量最低。初步证明PAL基因在不同品种的龙眼叶片中的表达量差异显著。
To obtain PAL cDNA sequence and its expression in 4 longan varieties’ leaves,the reverse transcription PCR,RACE and Real Time PCR technologies were used.The results showed that Dlpal cDNA completed sequence contained 2 532 base pairs and the Open Reading Frame（ORF）contained 2 101 base pairs.Dlpal codes 839 amino acids,the molecular weight and pI was predicted as 92.259 ku,7.38,respectively by DNAMAN software.The sequence is similar to PAL sequences of other plants.The expression profile showed that Dlpal in Lidongben’ was higher than other 3 varieties.There is remarkable difference of Dlpal expression in 4 varieties’.
出处
《热带作物学报》
CSCD
北大核心
2012年第1期79-83,共5页
Chinese Journal of Tropical Crops
基金
福建省自然科学基金项目(No.2011J01082)
