Abstract:A 901 bp fragment of tropomyosin gene (TPMS) was cloned from muscle of Litopenaeus vannamei using primers designed on the basis of homologous genes of other shrimps. A complete open reading frame of 852 bp coding a 32.8 kDa protein was found in the fragment. Results of RT-PCR indicated that the gene expressed in all examined tissues including heart, hepatopancreas, stomach, gills, intestine and muscle with different expressing levels. The strongest signal was found in muscle and the weakest signal was in gills. TPMS was sub cloned into pET 30a and expressed in E.coli BL21 (DE3). After induction of IPTG, a 38.2 kDa protein was largely expressed and formed inclusion bodies. Five hours inducing period and 0.05 mmol/L IPTG were the best conditions for recombinant expression. Recombinant expressed TPMS could be purified with His Bind Resin and the purity was higher than 90%.
