Cloning Of RRNA Repeating Gene Family Of Siniperca Chuatsi

The DNA sequences of rRNA repeating gene family of Tetraoden nigrovirdis were gained by bioinformatics technique of relative organism method. Based on the sequence three pairs of primers (P1/P2, P3/P4 and P5/P6) were designed to amplify three parts of the rRNA gene from genome DNA extracted from Siniperca chuatsi. Furthermore, the PCR products were cloned into pUCm-T cloning plasmid, and sequenced. The results from sequencing showed that the lengths of three cloned fragments of rRNA gene were corresponded to 3,321bp, 2,873bp and l,502bp, respectively. The integrated contig with three sequences was identified to 7,139bp, exactly, which separately comprised Siniperca chuatsi rRNA gene of 18S, ITS1, 5.8S. ITS2 and 28S with the length of l,837bp, 745bp, 158bp, 434bp and 3,073bp. These above sequences were submitted to GenBank, and the corresponding accession numbers were granted to AY452489. AY452490, AY452491, AY452492, AY452493, AY452494 and AY452495, respectively. Based on the analysis of nucleotide sequence homology in phylogenetic rRNA repeating gene family sequences from Siniperca chuatsi, human, Norway Rat, Tetraoden nigroviridis, oreochromis esculentus, cyprinus carpio and oncorhynchus mykiss (rainbow trout), the pairwise nucleotide sequence homology of 18S, 5.8S, 28S, ITS1 and ITS2 among 7 organisms were separately showed to be 91.2-98.6%, 95.0-100%, 87.2-92.8%, 6.8-62.9% and 11.9-64.0%. In terms of that, the phylogeny trees were constructed. The key material of gene targeting technique of multiple site for siniperca chuasti has been got in the experiment.