| 1. | According to the sequences of the conservative region of avain chlamydia momp genes reported by people abroad , one pair of primers were designed to amplify momp gene with pcr whose template was genome dna of this strain , then it was sequenced
其次，根据国外报道的禽衣原体momp基因两端保守序列设计合成一对引物，以提取的衣原体基因组dna为模板， pcr扩增、克隆了momp基因，并对其进行了测序。 |
| 2. | Momp gene is amplified by using pcr technologyfrom dna of l2 trachoma chlamydia . the pcr products are recombined with vectors of pmd18 - t . more over , the recombinant plasmids are colonged . the dna sequence analysis shows the insert fragment is momp dna
本实验采用pcr技术，从l _ 2型沙眼衣原体dna中扩增出momp基因，与pmd18 - t载体重组行t a克隆，并进行dna序列分析，证明所克隆基因为mompdna序列。 |
| 3. | For creating a reasonable displaying method , momp gene fragment and pbv221 are recombined oritationally they are transformed into e . coli jm109 ( de3 ) . a expressing protein was got with momp antigen . this study establishes the foundation for advance research of momp
设计一条较为合理的表达方案，将momp基因片段与pbv221定向重组转化大肠杆菌jm109 ( de _ 3 ) ，得到具有momp抗原性的表达蛋白，为研究momp的生物学功能以及研制沙眼衣原体基因工程亚单位疫苗奠定了基础。 |
| 4. | Lastly by using the technique of dot blot hybridization , the genome dna of chlamydia was detected with the probe of momp gene labeled with dig - 11 - dutp by using the way of random primer . the results showed the degree of sensitivity of the probe was 10 pg and other pathogens could not be detected by this probe . by comparing the diagnostic ways of nucleotide probe and fc , the technique of nucleotide probe were proved to have high sensitivity and speci fi city
最后，用地高辛随机引物法标记成momp基因核酸探针，斑点杂交检测衣原体基因组dna ，灵敏度可达10pg ，且不能检出其它病原体的核酸。将核酸探针法与补体结合反应法对衣原体感染的诊断进行比较，初步证明该探针具有较高的敏感性与较强的特异性。 |
| 5. | 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 ) 。 |
| 6. | 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 ) 。 |
| 7. | 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。 |
| 8. | 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。 |
