鳗源嗜水气单胞菌-溶血素基因的克隆及表达-水产学报.PDF

27 2 V ol 27 , No 2 2003 4 JOURNAL OF FISHERIE S OF CHINA Apr , 2003 : 1000- 0615( 2003) 02- 0124 - 07 - 龚 晖, 林天龙, 俞伏松, 董传甫, 陈日升, 杨金先, 陈振海 ( , 350003) : PCR , 1 M L316 - AHL316HEM , AHL 316HEM pGEM- T Easy Vector, p cDNA3 0 , PDLHPDLH ( Escherichia coli) DH5- , 4 - 1 1 28 10 HUmg , , - , PDLH - , : ; - ; ; : Q78 : A Cloning and expressing of the -hemolysin gene of Aeromonas hydrop hila strain isolated from Anguilla anguilla GONG Hui, LIN Tian- long, YU Fu- song, DONG Chuan-fu , CHEN R-i sheng, YANG Jin- xian, CHEN Zhen- hai (A nimal H usbandry and Veterinary Medicine Institute, Fuj ian Academy of Agricultural Sciences , Fuz hou 350003, China Abstract: With the technology of PCR, the completed - hemolysin gene AHL316HEM was obtained from the amplification of an Aeromonas hydrop hila strain ML316 ( isolated from diseased Anguilla anguilla The gene w as cloned into pGEM- T Easy Vector After analysis, the gene w as in serted into pcDNA 3 0 vector to con struct the recombinant plasmid PDLH The E . coli DH5w ith PDLH could lead to obvious - hemolysis plaque in blood- Agar The hemolysis activity of crude purif ied transf ormants extracellular products( ECP) w as 4 - 1 1 28 10 HU mg Furthermore, the transformant s ECP could be recognized by anti- serum which w as raised again st original bacterial strain The above results demonstrated that the - hemolysin gene of ML 316 had been cloned and the recombinant plasmid PDLH could express the product w ith native bio logical f unction The success of cloning and expressing the