ABSTRACT
<p><b>OBJECTIVE</b>To clone calmodulin (CaM) gene in Eleutherococcus senticosus, and study the effect of endophytic fungi on expression amount of CaM gene.</p><p><b>METHOD</b>The CaM full length cDNA sequence was cloned by rapid amplification of cDNA ends (RACE). The gene was analyzed and corresponding structure and functions were predicted by the bioinformatics methods. The expression amount of CaM gene affected of endophytic fungus P116-1a, P116-1b, P1094 and P312-1 was detected by RT-PCR.</p><p><b>RESULT</b>The full length of CaM cDNA was 856 bp containing an ORF of 450 bp that encoded a protein of 149 amino acids. The homologous of predicted protein was almost 100% with plants like Panax ginseng and Daucus carota. RT-PCR results showed that endophytic fungus improved CaM expression amount significantly (P<0.05). The highest expression amount of CaM occurred 90 d after reinoculated with endophytic fungi P1094, up to 2.96 times of the control.</p><p><b>CONCLUSION</b>The CaM gene of E. senticosus was successfully cloned for the first time. The results demonstrated that endophytic fungus of E. senticosus improved CaM expression amount significantly.</p>
Subject(s)
Calmodulin , Chemistry , Genetics , Metabolism , Cloning, Molecular , Eleutherococcus , Classification , Genetics , Metabolism , Microbiology , Endophytes , Physiology , Fungi , Physiology , Gene Expression Regulation, Plant , Molecular Sequence Data , Phylogeny , Plant Proteins , Chemistry , Genetics , MetabolismABSTRACT
<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>