嗜麦芽假单胞菌酪氨酸酶基因在类产碱假单胞菌中的转移与表达

类产碱假单胞菌 (Pseudomonaspseudoalcaligenes)是从盐城市农村粪池中采集的自然病死蝇蛆体内分离的具有显著杀蛆作用的细菌 ,野外使用易受阳光中紫外线的影响而失活。黑色素具有很强的抗辐射作用。将构建的含有嗜麦芽假单胞菌酪氨酸酶基因的质粒pWSY导入类产碱假单胞菌体内 ,使后者获得了稳定产生黑色素的能力。Southern杂交实验证实酪氨酸酶基因来源于嗜麦芽假单胞菌。SDS PAGE电泳显示该重组子体内额外表达了一分子量约为 1 8kD的蛋白 ,该蛋白很可能就是重组子表达的酪氨酸酶。经测定 ,重组子抗辐射作用明显增强 ,有效杀蛆时间显著延长 ,对畜、禽安全

TRANSFER AND EXPRESSION OF TYROSINASE GENE OF PSEUDOMONAS MALTOPHIIIA IN PSEUDOMONAS PSEUDOALCAIIGENES

Pseudomonas pseudoalcaligenes is a notably killing maggots bacterium,which was isolated from natural dead maggots in the manure pits in the countryside of Yancheng.It easily die from effect of ultra\|violet ray when it is exposed in the sun.Antiradiation effect of melanin is quite strong. Mel gene of P.maltophilia AT18 has been introduced into P.pseudoalcaligenes, and enabled it the ability of producing melanin steadily.Southern hybridization studies confirmed that the small fragment cloned in the P.pseudoalcaligenes comes from P.maltophilia DNA.SDS\|PAGE analysis also revealed that an additional protein of 18kD,which was equal to the size of the putative tyrosinase according to mel fragment,was exressed in the P.pseudoalcaligenes recombinant carrying the mel gene,The results of assay show that antiradiation effect of recombinant is quites strong,the effect time of killing maggats is longer than the recipient,the recombinant can't infect animals and fowls.