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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。

Peasants in one commune there secretly started parcelling out land, expecting death for it, but soon gained backing from a provincial leader and Deng ally, Wan Li.

一个公社的一些农民秘密地承包土地，甚至都做好了赴死的准备，但是不久他们得到了省领导和邓小平的助手，万里的支持。

Peasants in one commune there secretly started parcelling out land, expecting death for it, but soon gained backing from a provincial leader and Deng ally, Wan Li. Others gradually followed suit.

一个公社的农民将土地秘密分成小块耕种，他们预计到有可能因此丧生，但是他们的行动却很快得到了该省领导、邓的盟友万里的支持。

According to the results of the correlation matrix and partial correlation coefficient gained from polynomial linear regression, the relative contribution that the essential factor made to the number of tourist and tourism gross revenue of forest parks was analyzed.

结果表明，在兰州地区对森林公园客流量贡献最大的指标是人文景观、森林覆盖率和水体，对旅游总收入贡献最大的指标是人文景观，其次为森林覆盖率、旅游设施、水体，贡献最小的是山体。

The results point out that the values of kinetic parameters are altered with the addition of Ti and C, which change the crystallization type from difficult nucleation and easy growth pattern to easy nucleation and difficult growth pattern, and this difficult nucleation and easy growth pattern may be the main reason why fine and even microstructure is gained in Ti and C doped alloys.The influences of Nd content, B content, partial substitution of Nd by Pr on the microstructure and magnetic properties have also been researched.

文章从动力学理论的观点揭示了合金显微结构与合金晶化动力学特性之间的关系，指出Ti和C添加改变了Nd9.4Fe79.6B11合金晶相析出时的动力学参量值，使合金晶化时晶相的析出方式由难形核易长大型加热管转变为易形核难长大型，这种易形核而难长大的晶化动力学特征，是Ti和C添加合金获得细小均匀的纳米晶结构的根本原因。

In the authors opinion, one important reason why interests separation and political stability issues become more and more prominent in modern Chinese society is that over the past 26 years of reform and opening up to the outside world, a group of people who are named as parvenu gained the most interests from illegal and unjustifiable ways, the economic and political status of them are expanding continuously.

笔者认为，改革开放26年来形成的获利最大者群体中的那些不合法、不合理暴富者经济政治地位不断膨胀的趋势，是当代中国社会利益分化与政治稳定问题日益突出的一个重要原因。

My intimacy with the pasha and his unlimited confidence had gained me an introduction to them, and I had seen them above twenty times.

我和总督的密切关系以及他对我的忠诚的无限信任使我见过她们二十多次。'

Since the reforming and opening especially being new centurythe pawn broking have gained a good many development.

结论内容提要：典当业改革开放以来，特别是进入新世纪得到了很大的发展。

His peaceable methods have gained the world's respect; nothing more.

他的和平方案赢得世界的尊敬：毕竟，没有再好的办法了。

In 2008, Cai Wei who was working as a pedicab driver had already gained some accomplishments on ancient Chinese study.

早在2008年，那时还是三轮车夫的蔡伟就已经在古汉字研究上小有成就。

As she looked at Warrington's manly face, and dark, melancholy eyes, she had settled in her mind that he must have been the victim of an unhappy attachment.

每逢看到沃林顿那刚毅的脸，那乌黑、忧郁的眼睛，她便会相信，他一定作过不幸的爱情的受害者。

Maybe they'll disappear into a pothole.

也许他们将在壶穴里消失