Extraction of Total RNA from Horned-gall and the Cloning of ACTIN Gene Fragment

The extraction of total RNA from the horned-galls of Rhus chinensis is the basis of the researches on the molecular mechanism of gall formation.Due to high levels of polyphenol and polysaccharides,the modified CTAB method depositing with isopropanol was selected from different kinds of kits and other improved methods.It is a simple,economic and effective method for the extraction of total RNA,and the yield could get 60.16 μg/100 mg.Reverse transcription was conducted on the total RNA extracted by CTAB-isopropanol method,and a 934 bp cDNA sequence of ACTIN gene coding a protein of 311 amino acids was obtained.Homology comparison showed that it shared 95% nucleotide sequence homology with Mangifera indica and over 86% amino acid sequence similarity with actins in other species.The results laid basis for molecular researches of Rhus chinensis and other aphid induced galls.