枇杷Amyrin Synthase(AS)基因的5'RACE与3'RACE扩增及序列分析

5'RACE and 3'RACE of Amyrin Synthase Gene and Sequence Analysis in Eriobotrya japonica L.

以‘解放钟’枇杷(Eriobotrya japonica L.)叶片为材料,在前期获得保守区的基础上,采用RT-PCR结合RACE技术,扩增得到枇杷三萜合成途径中关键酶香树脂醇合成酶基因的5'和3'序列,并进行序列分析。结果表明,AS基因5'非编码区长96 bp;3'非编码区长321bp,在GenBank中更新的登录号为JX173279.2,结合已发表保守区序列,通过拼接,得到AS基因全长2 700 bp,含有一个2 283 bp的开放阅读框,编码760个氨基酸。生物信息学分析表明,该蛋白是定位于细胞核的蛋白,具有29个磷酸化位点,属于ISOPREN_C2_like超家族,含有Camelliol C synthase、squalene/oxidosqualene cyclases、Squalene cyclase多结构域,与苹果AS基因98%同源。

5＇cDNA and 3＇cDNA of AS （Amyrin systhase Gene）,which was an important enzyme of terpenes synthesis were cloned with RTPCR and RACE from Eriobotrya japonica L.leaves on the basis of the gene conserved region we obtained.And then,the sequence was analyzed.The results showed that：the full length of ej-AS mRNA （Updated GenBank accession number,JX173279.2）,about 1 775 bp,consisted of an Open Reading Frame of 1 239 bp,and 5＇and 3＇un-translated regions of 97 bp and 439 bp,respectively.The putative protein had 760 amino acids,and the identity to that of Malus domestica was 98％,and belonged to ISOPREN C2 like super family,contained Camelliol C synthase,squalene/oxidosqualene cyclases,Squalene cyclase multiple domains.