{"refrec":{"BRefID":131651,"RR":"<b>Dai, J.-G.; Xie, H.-W.; Jin, G.; Wang, W.-G.; Zhang, Y.; Guo, Y.</b> (2009). Preliminary study on high-level expression of tandem-arranged tachyplesin-encoding gene in <i>Bacillus subtilis</i> Wb800 and its antibacterial activity. <i>Mar. Biotechnol. 11(1)</i>: 109-117. <a href=\"https://dx.doi.org/10.1007/s10126-008-9125-6\" target=\"_blank\">https://dx.doi.org/10.1007/s10126-008-9125-6</a>","BEntID":125698,"PublicFlag":1,"CheckedFlag":1,"wosflag":1,"vabbflag":0,"RefStringPartII":". <i>Mar. Biotechnol. 11(1)</i>: 109-117. <a href=\"https://dx.doi.org/10.1007/s10126-008-9125-6\" target=\"_blank\">https://dx.doi.org/10.1007/s10126-008-9125-6</a>","DocTypID":8,"DocType":"Journal article","MarineFlag":1,"FreshFlag":0,"BrackishFlag":0,"TerrestrialFlag":0,"Authorstring":"Dai, J.-G.; Xie, H.-W.; Jin, G.; Wang, W.-G.; Zhang, Y.; Guo, Y.","OrigTitleTranslFlag":0,"Authorstringtrunc":"Dai, J.-G. <i>et al.</i>","Englishabstract":"To produce tachyplesin, an antimicrobial peptide, by a stable and efficient gene engineering approach, cDNAs containing single tachyplesin gene sequence (<i>tac</i>)<sup>1</sup> and tandem repeat of tachyplesin gene sequence (<i>tac2</i>) were respectively developed by annealing two synthesized complementary single-stranded DNAs and constructed into pSBPTQ shuttle vector under the control of the <i>SacB.p.s</i> promoter. The vectors containing the target gene sequence were then transformed into Bacillus subtilis WB800, respectively. Both expression of tac and <i>tac2</i> were induced by 2% sucrose. The fermentation supernatant was purified by regenerated cellulose membrane tubing (MWCO 2000) and the secreted TAC<sup>2</sup> and TAC2 were about 5 and 10 mg/l of supernatant, respectively. The antimicrobial activities of TAC and TAC2 were measured by the size of bacteriostatic circle of the fermentation supernatants against <i>Escherichia coli</i> K88. Ultrastructural alteration of <i>E. coli </i> K88 and <i>Salmonella typhimurium</i> was observed under scanning electron microscope and transmission electron microscopy. The results showed that in comparison with TAC, TAC2 was expressed at a higher level and also indicating strong antimicrobial activity both in vitro and in vivo. 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