Xing Zhaobin, Cao Lei, Chen Long, He Shan, Li Baocai, Zhu Jinli
College of Life Science, Hebei United University, Tangshan 063000, China.
Zhongguo Zhong Yao Za Zhi. 2012 Jan;37(2):172-5.
To clone and sequence the cDNA of squalene epoxidase gene in Eleutherococcus senticosus.
Total RNA of E. senticosus was extracted by the improved isothiocyanate method and reverse transcripted into cDNA. The primers were designed depending on the reported SE cDNA sequences of Panax ginseng. The SE cDNAs in E. senticosus was amplified using RT-PCR strategy.
Sequencing results showed two different cDNA fragments (SE1, SE2) with 1665, 1629 bp each ORF which encoded 554,542 amino acids, respectively. The identities of nucleotides and amino acids between SE1, SE2 were 91.49%, 92.55%. SE1, SE2 had the highest amino acids similarity to the SE1 of P. notoginseng, 93.45%, 94.87% respectively. SE1, SE2 both had a FAD binding domain. The deduced speculated amino acids of SE1, SE2 each had 2,4 membrane-spanning helices.
The two SE sequences in E. senticosus were firstly separated and reported, which has made foundation for E. senticosus secondary metabolite engineering researches.
克隆刺五加鲨烯环氧酶基因的cDNA并进行测序。
采用改良异硫氰酸胍法提取刺五加总RNA，并反转录成cDNA。根据已报道的人参鲨烯环氧酶cDNA序列设计引物，运用RT-PCR策略扩增刺五加中的鲨烯环氧酶cDNA。
测序结果显示有两个不同的cDNA片段（SE1、SE2），其开放阅读框分别为1665、1629 bp，分别编码554、542个氨基酸。SE1与SE2之间核苷酸和氨基酸的同一性分别为91.49%、92.55%。SE1、SE2与三七的SE1氨基酸相似性最高，分别为93.45%、94.87%。SE1、SE2均具有一个FAD结合结构域。推测的SE1、SE2氨基酸序列各有2个、4个跨膜螺旋。
首次分离并报道了刺五加中的两个鲨烯环氧酶序列，为刺五加次生代谢产物工程研究奠定了基础。
