Cloning,expression and adhesive characterization of the major adhesion gene in Aeromonas hydrophila isolated from Anguilla anguilla
1.Biotechnology Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003,China;
2.Luojiang Agriculture and Water Conservancy Bureau,Quanzhou 362011,China
Abstract:Aeromonas hydrophila strain ZN1 was isolated from European eel (Anguilla anguilla). Its major adhesin gene (Mah) was amplified by PCR and the DNA sequence analysis showed that Mah had a fulllenghth open reading frame (ORF) of 1074 nt, encoding a 357aa peptide, and which Nterminal 22 aminoacid was highly hydrophobic signal polypeptide. The homologous comparison proved that Mah had about 70% homology with the other Mah sequences from A. hydrophila. The amplified Mah was inserted into vector pET32a and transformed into E.coliDE3(BL21) for expression. The MahTrxA fusion protein was efficiently expressed in E.coliDE3(BL21) which were recognized by mouse antiZN1MOMP serum by ELISA and also Westernblotting to get the expected bands comparing with ZN1MOMP, which was containing major adhesin. Adhesive assay indicated that ZN1MOMP and MahTrxA fusion protein were able to inhibit the adhesive fuction of the bacteria and the rabbit antiMahTrxA serum could significantly block the ZN1 to adhere to EPC monolayers. So that, we could have the conclusion that the cloned gene represented the major adhesin gene of strain ZN1, and the gene products shared similar antigenicity with the natural major adhesion.
庄培德, 杨金先, 吴学敏, 龚晖, 林天龙. 鳗源嗜水气单胞菌主要粘附素基因克隆表达及产物粘附功能分析[J]. 中国人兽共患病学报, 2011, 27(2): 112-116.
ZHUANG Pei-De, YANG Jin-Xian, WU Xue-Min, GONG Hui, LIN Tian-Long. Cloning,expression and adhesive characterization of the major adhesion gene in Aeromonas hydrophila isolated from Anguilla anguilla. Chinese Journal of Zoonoses, 2011, 27(2): 112-116.
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