| The research on cyclin H (Pmcyclin H) is necessary for understanding the mechanisms involving ovarian developmental processes of giant tiger shrimp (Penaeus monodon). A high level prokaryotic expression of Pmcyclin H in E.coli BL21 (DE3) was set up, Pmcyclin H gene was cloned into expression vector pET-21a, which then was determined by double -endonuclease digestion and DNA sequencing. The recombinant vector was transformed into E.coli BL21 (DE3), and was induced to express fusion protein by 0.6 mmoL/L IPTG. The quality of expression product was identified by SDS-PAGE and Western blot; the recombinant protein was purified through Nichelating affinity chromatography, and the purified protein was identified by SDS-PAGE gel scan analysis and mass spectrometry. At 22%, both LA and YTGA medium could be used, Pmcyclin H protein was more abundantly expressed as an insoluble than soluble protein; at 37益, LA medium was better, and all the product was an insoluble protein. This study provides a fundamental condition supporting researches on structure, function and biological activity of cyclin H of P. monodon. |
