副结核

Paratuberculosis (MAP,Mycobacterium paratuberculosis) induce a series of characteristics as chronic hyperplastic enteritis andprogressing extenuation in ruminants also called Johne\'s disease.

副结核病是由副结核分枝杆菌即禽分枝杆菌副结核亚种Mycobacterium avium subsp。

This finding does not support a significant role for Mycobacterium avium subspecies paratuberculosis in the pathogenesis of Crohn's disease in the majority of patients.

这个发现也不支持鸟分支杆菌的亚型副结核分支杆菌是大多数克隆氏病患者的病因。

This finding does not support a significant role for Mycobacterium aium subspecies paratuberculosis in the pathogenesis of Crohn's disease in the majority of patients.

这个发现也不支持鸟分支杆菌的亚型副结核分支杆菌是大多数克隆氏病患者的病因。

Bovine paratuberculosis is caused by Mycobacterium paratuberculosis and it is a kind ofchronic wasting disease in ruminants globally, which is called Johnes disease.

副结核病是由副结核分枝杆菌引起的一种全球性以反刍动物为主的慢性、消耗性传染病，也称Johnes。

The expression protein was analyzed by using the Western-blot, which proved that it had theantigenic activity of Mycobacterium paratuberculosis.

经免疫印迹分析证实，该融合蛋白具有副结核分枝杆菌抗原性。

Studies show the M. paratuberculosis secretes immunological competent protein in short termculture and opposites in long term culture.

研究表明，在短期培养过程中，副结核分枝杆菌分泌具有免疫活性的蛋白质，而在长期培养则相反。

In this experiment, we screen the major protective antigen gene-SOD gene of M. paratuberculosisin order to study the sensitive, specific diagnostic reagent and prophylaxis preparation, especially theDNA vaccine. The SOD gene was amplified from Mycobacterium paratuberculosis C-2 chromosomalDNA by using the PCR technique and cloned into pMD18-T Vector System. We gained a SOD gene of624bp.The recombinant clone was identified byα-complementarity, enzyme digestion and PCRidentification. The result indicated that the recombinant plasmid pMD18-T-SOD was successfullyconstructed. Moreover, through sequential determination and DNASTAR analysis between the clonedSOD gene of M. paratuberculosis C-2 and that of the M.paratuberculosis K-10 strain, the sequentialhomogeneity reached 99%, and the amino acid homogeneity reached 99.5%. The preceding analysisindicated that the SOD gene was very conservative in M. paratuberculosis.

为了研制副结核病敏感、特异的诊断试剂和新型、高效的预防制剂，尤其是DNA疫苗，本研究筛选了M.paratuberculosis主要保护性抗原SOD基因，以M.paratuberculosis C-2染色体DNA为模板，以SOD基因的特异性引物进行PCR扩增，获得了624bp的SOD基因，通过T-A克隆技术，将PCR产物克隆至pMD18-T Vector中，以质粒大小、酶切分析、PCR扩增及序列分析鉴定重组克隆，成功地构建出克隆质粒pMD18-T-SOD，序列测定及DNASTAR分析表明，所获得的M.paratuberculosis C-2 SOD基因与Gen Bank中M.paratuberculosis K-10 SOD基因的大小完全一致，两者核苷酸序列的同源性为99％，氨基酸序列的同源性为99.5％，表明该基因在副结核分枝杆菌中是高度保守的。

In order to develop an indirect ELISA for diagnosing bovine paratuberculosis, the DNA fragmentsof map086 2and map2154c were amplified from the genome DNA Mycobacterium avium Subspeciesparatuberculosis K10. Then the DNA fragments of map0862 and map2154c were spliced byoverlapping extension, yielding a fusion gene map0

为了建立一种有效的牛副结核病间接ELISA方法，本研究设计了两对引物，从副结核分枝杆菌P_(18)株的基因组中PCR扩增出两个目的基因map0862和map2154c、，采用重叠延伸剪接技术(splice by overlapping extension，SOE)获得融合基因map0

It also can be used as alternative antigen of newgeneration vaccine.In this experiment,we screen the major protective antigen hsp65 gene of MAP in order to developnew vaccine especially the DNA vaccine for the prevention of paratuberculosis disease.The hsp65 genewas amplified from MAP C-2 chromosomal DNA by using the PCR technique.We gained a hsp65 gene of 1 626bp.Then PCR product was cloned into pGEM-T vector by T-A clone technique and therecombinant clone was identified by plasmid size,enzyme digestion and PCR identification.The cloneplasmid of pGEM-T- hsp65 was successfully constructed.The nucleotide sequence and deduced aminoacid sequence ofclone gene was analyzed by DNASTAR software.The result indicated that the size ofhsp65 gene consist with M.paratuberculosis K-10 strain in GenBank and the sequential homogeneityreached 99.1%,the amino acid homogeneity reached 99.3%.The preceding analysis indicated that thehsp65 gene was very conservative in M.paratuberculosis.

为了研发预防副结核病的新型疫苗尤其是DNA疫苗及相关蛋白功能，本研究选择了MAP的主要保护性抗原Hsp65蛋白，以副结核分枝杆菌C-2株染色体DNA为模板，以hsp65基因的特异性引物进行PCR扩增，获得了1 626bp的hsp65基因，通过T-A克隆技术，将PCR产物克隆至pGEM-T Vector中，以质粒大小、酶切分析、PCR扩增及序列分析鉴定重组克隆，成功地构建出克隆质粒pGEM-T-hsp65，以DNASTAR软件分析了所克隆基因的核苷酸序列和推导的氨基酸序列，结果表明，所获得的hsp65基因与GenBank中MAPK-10株该基因核苷酸大小完全一致，两者核苷酸序列的同源性为99.1％，氨基酸序列的同源性为99.3％，表明该基因在副结核分枝杆菌中高度保守。

The plant Quarantine Lab had intercepted more than 200 quarantine pests such as : Tilletia indica , Tilletia controversa , Bursaphelenchus xylophilus , sorghum halepense , Acanthoscelides obtectus , Sternochetes mangiferae , Bactrocera dorsalis , Coptoermes curvignathus for nearly 5000 batches ; The Animal Quanantine Lab had intercepted about 20 cow which had some pathogens evidence of infectious disease such as : bovine infectiouse rhinotracheitis , leukaemia bovum , paratuberculosis and so on ; The food microorganism lab had detected over 300 unqualified products such as Salm-Sury , Listeria monocytogenes , Mycobacterium paratuberculosis etc .

多年来，分中心各实验室切实履行严格把关的职责，植物检疫共截获大豆疫霉病菌、小麦印度腥黑穗病菌、小麦矮腥黑穗病菌、松材线虫、假高梁、菜豆象、芒果果核象甲、桔小实蝇、大家白蚁等国家禁止进境的危险性有害生物220 多种近10000 批次；动物检疫发现感染牛传染性鼻气管炎、牛白血病、副结核等传染病的疫牛20 多头；食品微生物检测共检测沙门、单增、副结核的不合格商品300 多批；分子生物学实验室为国内企业出具非转基因证书和不含有牛羊源成分证书50 余份，使相关的产品能够顺利出口到欧盟、韩国和日本等国。

The split between the two groups can hardly be papered over.

这两个团体间的分歧难以掩饰。

This approach not only encourages a greater number of responses, but minimizes the likelihood of stale groupthink.

这种做法不仅鼓励了更多的反应，而且减少跟风的可能性。