Second , the omps of aeromonas hydrophila ba3 were extracted with four experimental ways , namely ultrasonic break , ultrasonic - ep ( edta and pmsf ) break , abrasive break and abrasive - ep break . the best sds - page profile was observed when the omps were extracted by ultraisonic - ep break 分别用超声波法、超声波加ep ( edta和pmsf )法、研磨法和研磨加ep法四种不同方法提取了嗜水气单胞菌ba3株的外膜蛋白。
The major outer membrane proteins ( momps ) of an aeromonas hydrophila named ahl316 which isolated from diseased eel and the referential strain ahtps - 30 were purified and used for preparing immunostimulating complexs ( momp - iscoms ) . the immunotrial was carried out by injecting european eel peritoneally with 20ug of momp - iscoms per eel 本研究选取鳗源嗜水气单胞菌l316和参考菌株嗜水气单胞菌tps - 30 ，分别制备了这两株菌主要外膜蛋白免疫刺激复合物( momp - iscoms ) 。
The results demonstrated that the momps were protective antigens and the momp - iscoms of aeromonas hydrophila could induce the host to mount satisfied immunity . a pair of primers were designed according to the published nucleotide sequence of a putative outer membrane protein gene ( omp ) of aeromonas hydrophila . with the specific primers , a target fragment about 1 . 1kb was amplified from aeromonas hydrophila l316 via pcr . the target fragment was inserted into the linearized pgem - t easy vector 根据已发表嗜水气单胞菌的外膜蛋白基因omp的核苷酸序列设计引物，利用pcr技术，扩增、克隆了嗜水气单胞菌l316的主要外膜蛋白基因( momp ) ，经t a克隆，插入到pgem - t系列载体上，测序分析结果表明momp基因最长的开放阅读框( orf )为1035nt ，编码由344个氨基酸组成，分子量为36kda的主要外膜蛋白质( momp ) 。
Proteins were identified with peptide mass fingerprinting ( pmf ) . firstly , the first molecular anatomic profile of aeromonas hydrophila was established , 40 proteins were identitied from analysis of the pmf of l24 proteins . of the 40 proteins , 17 were from the source of aeromons hydrophila genes / proteins 实验首先初步构建了一张嗜水气单胞菌总蛋白质的分子解剖图谱，通过对124个蛋白质的肽质量指纹图谱分析，鉴定了40个蛋白质，其中17个为嗜水气单胞菌的蛋白。
We determine the growth process and mutual relationship of three bacteria , i . e . photosynthetic bacteria , e . coli ki2 and etec 987p . relationship between pathogenic bacteria in water , aeromonas hydrophija , and phoiosynthetic bacteria was studied emphatically . the result showed that when inoculating quantity of phoiosynthetic bacteria was one fifth of that of aeromonas hydrophila , the growth of latter bacteria could be inhibited distinctly 其次，研究了混合生长体系的细菌色谱行为，实时对混合培养体系中每一种细菌的生长情况作出定量和定性的分析，确定了光合细菌、 e . colik12和etec987p三种细菌混合生长的过程及相互竞争与抑制的作用关系；针对水体中病原菌? ?嗜水气单胞菌，重点考察了它与光合细菌的作用关系，结果表明光合细菌的接种量为气单胞菌的1 5时，就能够对气单胞菌有明显的抑制效果。
Make all these together , it proved that the cloned gene represented the major outer membrane protein gene of ahl316 , and the expressed gene products shared identical antigenicity with the natural main outer membrane protein . the studies on preparation and application of momp - iscoms of ah l316 provided a new approach to fish vaccinology . the successfully cloning and expressing the major outer membrane protein gene of ah l316 made it possible to describe this gene ' s function under a single factor level , and also provided technical support for developing an advanced gene engineering vaccine and subunit vaccine against aeromonas hydrophila 鳗源嗜水气单胞菌l316主要外膜蛋白免疫刺激复合物的制备与应用研究，对研制鱼类疫苗学问题进行了新的初步探索；成功地克隆和表达嗜水气单胞菌l316主要外膜蛋白基因为在单因子水平上研究嗜水气单胞菌外膜蛋白的作用和免疫功能以及制备嗜水气单胞菌基因工程疫苗和亚单位疫苗奠定技术基础。
The homologious comparison proved the cloned gene had 96 % homology to the sequence of the omp gene , and the alignment of the amino acid sequence was 98 % . the recombinant plasmid was constructed with the target gene and the expressing vector pgex - 4t - l and then was transformed into e . coli bl21 ( de3 ) the fusion protein was expressed under the iptg inducing condition , and exhibited about 62kda in size , very close to the predicted molecular weight of gst - momp . furthermore , the fusion protein was specifically recognized by anti - serum which raised against the major outer membrane protein of ahl316 Sds - page电泳分析显示诱导表达的基因产物分子量约为62kda ，与预测的gst -外膜蛋白重组融合蛋白的分子量极为相似， western - blot进一步证实，表达产物能被嗜水气单胞菌l316主要外膜蛋白特异性抗血清所识别，产生明显的染色条带，说明所表达的基因产物与天然的外膜蛋白抗原性一致。
