cDNA fragment encoding complete sequence of silver carp(Hypophthalmichthys molitrix) microcystin-deto-xifizyme was subcloned to construct a fusing gene expression vector pEGFP-N1-sGST,and a dicistronic expression vector pIRES2-EGFP-sGST.Using lipofectin method,the fusing gene expression vector pEGFP-N1-sGST was transfected into Hela cells.The emission of fluorescence was observed after 60 hours.Through microinjection,the dicistronic expression vector pIRES2-EGFP-sGST was delivered into embryos of zebrafish(Danio rerio),and the silver carp microcystin-detoxifizyme transgenic zebrafish was obtained.The success of the two transgenic models makes it possible to further analyze the cis-regulatory elements of the microcystin-detoxifizyme gene,and to develop a strain of transgenic planktivorous fish,used as a high-efficient biodetoxicator for microcystin in lakes and reservoirs.
[1] Krienitz L,Ballot A,Kotut K,et al.2003.Contribution of hot spring cyanobacteria to the misterious deaths of lesser Flamingos at Lake Bogoria,Kenya[J].FEMS Microbiol Ecol,43:141-148.
[2] Codd G A.1995.Cynobacterial toxins:occurrence,properties and biological significance[J].Water Sci Technol,32:149-156.
[3] Dawson R M.1998.The toxicology of microcystins[J].Toxicon,36:953-962.
[4] Carmichael W W.2001.Health effects of toxin-producing cyanobacteria:"the CyanoHABs"[J].Hum Ecol Risk Assess,7:1393-1407.
[5] Yu S J.1995.Primary prevention of hepatocelluar carcinoma[J].J Gastroenterol Hepatol,10:674-682.
[6] Pouria S,Andrade A D,Barbosa J,et al.1998.Fatal microcystin intoxication in haemodialysis unit in Caruaru,Brazil[J].Lancet,352:21-26.
[7] Chorus I,Falconer I R,Salas H J,et al.2000.Health risks caused by freshwater cyanobacteria in recreational waters[J].J Toxicol Environ Health,3:323-347.
[8] Hitzfeld B C,Hoger S J,Dietrich R.2000.Cyanobacterial toxins:removal during drinking water treatment,and human risk assessment[J].Environ Health Perspect,10:113-122.
[10] Xie P,Liu J K.2001.Practical.success ofbiomanipulation using filter-feeding Fish to control cyanobacteria blooms:a synthesis of decades of research and application in a subtropical hypereutrophic lake[J].Scientific World Journal,1:337-356.
[12] Datta S,Jana B B.1998.Control of bloom in a tropical lake:grazing efficiency of some herbivorous fishes[J].J Fish Biol,53(1):12-24.
[13] Pflugmacher S,Wiegand C,Oberemm A et al.1998.Identification of an enzymatically formed glutathione conjugate of the cyanobacterial hepatotoxin microcystin-LR:the first step of detoxication[J].Biochim Bioph Acta,1425:527-533.
[15] Zhu Z,Xu K,Li G,et al.1986.Biological effects of human growth hormone gene microinjected into the fertilized eggs of loach[J].Kexue Tongbao,31(11):988-990.
[17] Du S J,Gong Z,Fletcher G L,et al.1992.Growth enhancement intransgenic Atlantic salmon by the use of an"all fish" chimeric growth hormone gene construct[J].Bio/Technology,10:176-181.
[18] Maclean N,Rahman M A,Sohm F,et al.2002.Transgenic tilapia and the tilapia genome[J].Gene,295:265-277.
[19] Martinez R,uncal J J,Zaldivar C,et al.2000.Growth Efficiency in Transgenic Tilapia (Oreochromis sp.) Carrying a Single Copy of an Homologous cDNA Growth Hormone[J].Biochem Biophys Res Commun,267:466-472.
[20] Gao Y,Yue W,Zhang P,et al.2005.Spindlinl,a novel nuclear protein with a role in the transformation of NIH3T3cells[J].Biochem Biophys Res Commun,335(2):343-50.
[23] Tencalla F,Dietrich D.1997.Biochemical characterization of microcystin toxincity in rainbow trout (Oncorhinchus mykiss)[J].Toxicon,35(4):583-595.
[24] Wiegand C,Pfugmacher S,Giese M,et al.2000.Uptake,Toxicity,and Effects on Detoxication Enzymes of Atrazine and Trifluoroacetate in Embryos of Zebrafish[J].Ecotoxicol Environ Saf,45:122-131.
[25] Best J H,Eddy F B,Codd G A.2003.Effects of Microcystis cells,cell extracts and lipopolysaccharide on drinking and liver function in rainbow trout Oncorhynchus mykiss Walbaum[J].Aquat Toxicol,64:419-426.