Ural Licorice Root Cdna Library Construction And Est Analysis

Radix Glycyrrhizae is a traditional medicine which is wild used in clinicalpractice. Now the wild resources of Glycyrrhiza uralensis have been reducedgradually, and the cultivated licorice's quality isn't stabile. Gene is the innerfactor of the herb's quality. The study on the gene characters could show themechanism of the biosynthesis of components. To face this actuality, it isnecessary to search for the genes which adjust and control the process of effectualcomponents. As the basis of gene research, constructing a eDNA library in specialtime and special organs becomes a more convenient way to gene's commentaryand filtration. Therefore, a cDNA library is necessary to gene expression andquality explanation.Chosen the root tissues of Glycyrrhiza uralensis, we constructed a cDNAlibrary using the method of Molecule Biology, Bio-informatics, Plant physiologyetc. Then we did EST analysis and full-length gene analysis to study the genecharacters of Glycyrrhiza uralensis. The important conclusions are as follows:(1) Constructed the methods of RNA isolation and eDNA syethsis.Total RNA of Glycyrrhiza uralensis root and leaf tissue was isolated using themethod of lithium chloride sedimentation. And cDNA was obtained using themRNA isolating kit and cDNA syethesis kit of TaKaRa. The quality of RNA andcDNA was checked by UV and agarose gel electrophoresis.(2) Construct a eDNA library of Glycyrrhiza uralensis root tissue.A cDNA plasmid library of Glycyrrhiza uralensis has been constructed andthe capacity of the original library was 1.15×10~7 with a recombinant rate of 98.2%. The capacity of the amplified library was 1.41×10~9 with a recombinant rate of95.3%. The inserted eDNA fragments ranged from 0.5kb to 4.8kb, mostly from1.0kb to 2.5kb.(3) Analyzing the characters of Glycyrrhiza uralensis ESTs.We obtained 126 ESTs by 5' end and 3' end from random sequencing, theresult of EST clustering indicated that 70 gene clusters were obtained. Theunigenes ranged from 255bp to 3110bp. The results of functional definitionshowed that total 45 genes as genes of known function related to 86 ESTs. Andtotal 5 genes as hypothetical protein related to 13 ESTs. Then 8 genes asunidentified related 10 ESTs while 12 genes as none salience homological related17 ESTs. The mostly homological proteins were from leguminous plants,Arabidopsis thaliana, Oryza sativa, and so on. The proteins were nearly related toresisted enzymes and some deoxidize enzymes, hydrolyze enzymes and induced...