Previously, we found that expression by translational fusion of the polyhedrin (Polh)-green fluorescence protein (GFP) led to the formation of granular structures and these fluorescent granules were easily precipitated by high-speed centrifugation. Here, we developed an easy, fast, and mass purification system using this baculovirus expression system (BES). An enhanced GFP (EGFP) fused with Polh gene at the N-terminus including an adaptor and enterokinase (EK) site between Polh and EGFP was expressed in Sf9 cells. The cells infected by AcPolhEKA-EGFP produced fluorescent granules. The EGFP fusion protein was purified from granule-containing cells according to three steps; cell harvest, sonication and EK digestion. Through the final enterokinase digestion, EGFP was presented mainly in the supernatant (93.3%) and the supernatant also showed a pure EGFP band. These results suggest that the combined procedure of Polh fusion expression and enterokinase digestion can used for the rapid and easy purification of other proteins.

Sclerodermus harmandi populations were collected from 6 Korean regions (Seoul, Gangwon, Chungcheong, Jeolla, Gyeonasang and Jeju provinces), 1 Chinese region (Nanjing) and 1 Japanese region (Tokyo). Above all was identified as the same species based on morphological characteristics. In present study, we compared the intra-specific characteristics of regional isolated populations of S. harmandi based on nucleotide and amino acid sequences of variant genomic and mitochondrial loci: the full length of internal transcribed spacer 1 and 2 (ITS1 & ITS2), the complete cytochrome oxidase subunit I and II (COI & COII) and tRNA genes for aspartate and lysine and 5'end of ATPase 8 genes. There were no significant differences among the analysed 9 populations regardless of regional segregation. The nucleotide and translated amino acid sequences similarities were significantly high, despite some sequence differences were found in same loci. This result support the report based on the morphological characteristics. However, we are conducting further study to identify whether sequence differences in specific loci are the characteristics of individual level in population or population level and to analyse the full sequences of mitochondrial DNA by regional segregation.