Cloning and sequence analysis on cDNA of squalene epoxidase gene in Eleutherococcus senticosus / 中国中药杂志

ABSTRACT

<p><b>OBJECTIVE</b>To clone and sequence the cDNA of squalene epoxidase gene in Eleutherococcus senticosus.</p><p><b>METHOD</b>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.</p><p><b>RESULT</b>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.</p><p><b>CONCLUSION</b>The two SE sequences in E. senticosus were firstly separated and reported, which has made foundation for E. senticosus secondary metabolite engineering researches.</p>