摘要
根据NCBI数据库中已知的蔷薇科植物花青素合成酶基因序列设计特异引物,从本实验室已经构建好的木奈成熟果实均一化全长cDNA文库中筛选分离得到了一个编码花青素合成酶的cDNA:基因命名为PsANS,登录号:JN560957。该cDNA长1 478 bp,包含137 bp 5'非编码区,267 bp 3'非编码区,开放阅读框长度为1 074 bp。PsANS编码358个氨基酸,分子量为40.41 ku,理论等电点为5.35。经BLAST分析对比发现,PsANS氨基酸序列与甜樱桃、苹果和草莓的ANS氨基酸同源性都很高,分别为98%、93%和89%。将PsANS亚克隆到原核表达载体pGEX-6P-1和pET-32a上,在大肠杆菌BL21中经1mmol/L ITPG诱导表达获得了木奈PsANS融合表达蛋白,其分子量大小与预期的一致,进一步确认本试验克隆得到的PsANS为木奈花青素合成酶基因。
A cDNA encoded anthocyanidin synthase（ANS）was separated from a normalization and full-length cDNA library prepared previously with the ripe fruit of ＇oil Lai＇（Prunus salicina）by gene special primers designed from ANS genes of other Rosaceae plants. The eDNA named PsANS was 1 478 hp in full length and encoded a protein with 358 amino acids. The deduced amino acid sequence shares 98%, 93% and 89% identity with those A NS from sweet cherry, apple and strawberry, respectively. Prokaryotic expression was conducted with the constructed vectors used PsANS and prokaryotic expression vector（pGEX-6P-1 and pET-32a）in E.coli BL21. The obtained fusion protein had a molecular weight consistent with that expected, and it further confirmed that the PsANS obtained by this experiment encoded ANS of ＇oil Lai＇（Prunus salicina）.
出处
《热带作物学报》
CSCD
2011年第11期2088-2093,共6页
Chinese Journal of Tropical Crops
基金
国家科技支撑计划(No.2007BAD07B00)
校科技发展金(No.2009068)
福建省教育厅基金(No.JA10108)
作者简介
姜翠翠．女．博士研究生。研究方向：果树生物技术。
通讯作者：潘东明，E—mail：pdm666@126．com。
