关键词：
磷脂酰乙醇胺结合蛋白;原核表达;低温诱导;雪莲
摘 要：
根据雪莲PEBP基因序列设计特异性引物,利用实验室构建的Teasy-PEBP质粒为模板,扩增PEBP基因,与表达载体pET-30a(+)连接转化BL21(DE3),PCR和酶切鉴定筛选阳性克隆,测序分析。经IPTG诱导后SDS-PAGE检测,结果表明:PEBP得到大量表达,表达量占菌体总蛋白的26.8%,融合蛋白的相对分子量大约是28KD,与理论值相符,为进一步研究该蛋白的理化性质奠定了基础。
译 名：
Construction and Expression of Prokaryotic Vehicle Saussurea involucrate Kar.& Kir.PEBP
作 者：
ZHANG Xue-jun1,2,AI Xiu-lian1,LU Xiu-juan2,LUO Ming2,WANG Bo1 (1.Institute of Microbiology,Xinjiang Academy of Agricultural Sciences,Urumqi 830091,China;2.College of Agronomy,Xinjiang Agricultural University,Urumqi 830052,China)
关键词：
PEBP;Prokaryotic expression;cold-induced;Saussurea involucrate Kar.& Kir.
摘 要：
A pair of specific primers was designed and synthesized according to Saussurea involucrate Kar.& Kir.PEBP gene sequence.Using Teasy-PEBP plasmid as a template,the PEBP gene was amplified by PCR and inserted into Prokaryotic expression vechicle pET-30a(+),and transferred into the host bacteria BL21(DE3).The recombined plasmid was identified by PCR and zymosection analysis.The positive clones were selected and sequenced.After induced by IPTG,the PEBP was expressed effectively by SDS-PAGE analysis.The results showed that the fusion protein relative molecular weight was 28KD that was consistent with the theoretical value.Its expression product was 26.8% in the total proteins.It laid the foundation of the further study on the physical and chemical characters of the PEBP.
