摘要
从牡丹‘洛阳红’(Paeonia suffruticosa‘Luo Yang Hong’)花瓣中提取总RNA,根据GenBank上牡丹ACC合成酶基因PsACS的序列设计引物,通过RT-PCR获得牡丹PsACS1基因序列,包含一个1 479 bp的开放阅读框,编码492个氨基酸。将PsACS1基因cDNA片段与pET28a(+)构建原核表达载体pET-ACS1,转化大肠杆菌E.coli BL21(DE3)。0.4 mmol/LIPTG诱导3 h后,在预期的蛋白分子量55 kD处出现1条表达加强的蛋白条带。为进一步目的蛋白的纯化和鉴定提供试验基础。
Total RNA was isolated from the cut flower of tree peony ＇Luo Yang Hong＇.A pair of specific primers were designed according to ACC synthase ACS gene sequence of Paeonia suffruticosa recorded in GenBank.A cDNA fragment of PsACS gene including an ORF（1 479 bp）was amplified by RT-PCR,encoding 492 amino acids.The recombinant prokaryotic expression vector pET-ACS1 was constructed using vector pET-28a（＋）and transformed into E.coli DH5α.SDS-PAGE of PET-ACS1 protein induced by 0.4 mmol/L IPTG for 3 h showed that a strengthened protein band in expectant 55 kD protein marker.These results could provide a foundation for further purifying and identifying objective protein.
出处
《生物技术通报》
CAS
CSCD
北大核心
2011年第10期97-100,共4页
Biotechnology Bulletin
基金
国家自然科学基金项目(30972030)
关键词
牡丹
ACC合成酶基因
克隆
原核表达
Paeonia suffruticosa ACC synthase gene Cloning Prokaryotic expression
