Preliminary Studies On Cloning Of MicroRNAs From Salt Stressed Oryza Sativa Seedling

MicroRNAs (miRNAs) have now been accepted as playing the very important role during organism development and metabolizing courses by regulation of gene expression. Cloning of miRNAs is the basal work in study of expression and function of miRNA genes. So far, most plant miRNAs studies have focused mainly on model plants, such as Arabidopsis thaliana and Oryza sativa. Rice is one of the most important food crops and can provide more genomic data for analyzing gene structure and function. In this research, we tried to clon miRNAs from salt stressed rice seedlings after ameliorating RNA islation procedure. The following results were obtained:1. A new and efficient method for isolation of plant RNAs was developed by adding bentonite into extraction buffer in order to get rid of protein and restrain RNase. The result of argarose gel electrophoresis of extracted RNA from Jatropha curcas leaves showed that the new method was most efficient compared with TRIZOL reagen, SDS-KAc solution and Guanidine isothiocyanate solution. And successful RT-PCR of RNA showed that the RNAs isolate with this method were in good integrity and purity, and can meet the needs of most molecular biological experiments.2. The efficient and economical method for isolation of small RNAs was designed and developed based on the characteristic of Mg²⁺ capable to deposit efficaciously small nucleic acids using young corn plants (Zea may L). By optimizing parameters, such as Mg²⁺ concentration, sedimentation time and...