同源物

The nucleotide and amino acid homology of the 5 isolates is higher than 94% and 96% respectively.

PRSV 5个分离物核苷酸序列的同源性在94%以上，氨基酸序列的同源性在96%以上。

The novel primers were evaluated by detecting BTV serotypes 1, 3, 5, 8, 10, 11, 21 and 22. The specificity of the primers was estimated by comparing to gene sequences of viruses published in GenBank, and further assessed by detecting BTV serotype 1-12 and Epizootic hemorrhagic disease virus serotype 1-4. The sensitivity and repeatability of PCR with the novel primers were evaluated by successfully detecting the recombinant plasmid pGEM-T121 containing the diagnosed nucleotide sequence.

Blast软件分析引物与GenBank发表的病毒基因的同源性，显示引物特异性较好；利用8个不同血清型BTV标准毒株进行RT-PCR检测，证实该引物能有效检测不同血清型BTV；通过BTV1-12和EHDV1-4进行检测，证实该引物具有较好的检测特异性；通过制备的含特检序列的质粒标准品P121进行检测，显示该引物检测的敏感性为105 copies，重复检测CV%为0；对模拟血清样品进行检测，结果均为阳性。

For example, we have conducted the sequence screening on the binding sites of the interface of Barnase/Barstar complex and found that our DGA method can not only determine which position is more conservative and more important, but also redesign the binding sites rationally. The random sequence design for α,β,α/β and α+β four folding motifs have also been practiced and revealed that there are both homology evolution and heterology evolution of protein sequences compatible the same backbone motif.

例如，通过对典型的Barnase／Barstar复合物界面研究发现，它可以确定界面结合位点的重要性和保守性，并对结合位点进行合理的序列重新设计；通过对α、β、α/β、α+β等四种折叠模式骨架的随机序列设计，发现在蛋白质进化过程中，对同一种结构模式同时存在序列的同源进化和非同源进化这两种趋势。

In order to compare the homology of the gene encoding VP2 between MDPV-Q and MDPV which was registered in the GenBank, also in order to find out changes of the VP2 gene and the immuno-genicity between the wild-strain MDPV-Q and the attenuated strain MDPV-26 which derived by continued passage of virulent wild-type in muscovy duck embryo, a pair of primer (LHMP7/LHMP8) was designed. The upper one LHMP7 and the lower one LHMP8 corresponded to the MDPV-specific nucleotides sequence 2885-2900 and 4618-4604 respectively, according to the MDPV nucleotide sequence databases. The length of the sequence embraced by the primers was 1734bp.

为了获取这一基因片段与国外分离株进行同源性比较，同时也为了了解MDPV强、弱毒株VP2基因之间的异同关系，及其免疫原性的变化规律，通过DNA重组技术，设计了一对引物LHMP7／LHMP8，该对引物选取位于2885～2900及4618～4604的两段序列，跨幅为1734bp，并在这两条引物中分别加入两种限制性核酸内切酶SacⅡ和KpnⅠ的酶切位点，分别对MDPV-Q和由该株病毒经人工致弱后得到的MDPV-26株进行PCR扩增，并将PCR产物克隆到pMD18-T载体上，分别得到二个重组子：pMD18-T—M-Q VP2和pMD18-T-M-26 VP2。

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。

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 predicated gene products of amrA, amrB showed high similarities with the typical response regulator, such as afsQ1, mtrB and ompR, about 40%-62%.

对它们的氨基酸序列分析表明：amkA和amkB与典型的组氨酸激酶，包括afsQ2，mtrB以及phoR，有较好的同源性，大约为30％-44％；amrA和amrB与典型的反应调节物，包括afsQ1，mtrA和ompR等，同源性很高，大约为40％-62％。

Pneumoniae FH strain was cloned and the sequence was analysed by M13 DNA sequencing method. Comparing the PCR product sequcence with MP M-129 strain P1 gene, we found that there are 4 bases different. This may result from the different MP DNA templates. The maximum homology is 98.8%. The result confirmed the fidelity and specificity of the amplified target DNA segment by PCP, and suggested that two categories of MP P1 gene still exist a few differences even in the conservation region. The cloning MP DNA segment was labelled by random hexanucleotide priming, after hybridization, the probe detection was completed using an anti-digoxigenin antibody alkaline phosphatase conjugate, and the substrates 5-bromo-4-chloro-3-indolyl phosphate and nitro blue tetrazolium. This hybridization system is much superior to the radioactive probe hybridization, because it is safe, easy to handle and has no limitation of decay time. The time required for colormetic detection is also much less than the corresponding autoradiographic exposure time needed to achieve similar detection limits with 32P-labelled probes. The Dig-probes could be used repeatedly, and this made them not only much convenient to use, but also lower the cost, and worthwhile to be used popularly.

将PCR产物进行重组，并将阳性重组质粒，应用M13测序系统对产物进行DNA序列分析，并与MPM-129株P1基因核苷酸进行同源性比较，发现有4个位置的碱基发生了变化，其同源性为98.8％，证实了PCR所扩增DNA片段的准确性和特异性，同时也证实了不同MP组型的P1基因即使在保守区也存在着一定的差异，将克隆的目的DNA片段用异羟基洋地黄毒苷配基用随机引物法标记制备MP DNA探针，杂交后用碱性磷酸酶标记的抗Dig多克隆抗体与杂交体反应，再用BCIP和NBT呈色，制备MP DNA探针，鉴定所扩增片段的特异性，与同位素探针比较，Dig探针不受半衰期限制，可反复使用，而且价格低廉，值得推广使用。

Rapid amplification of cDNA end:Retroverse transcription product of total RNA extracted from normal porcine tissue was used as the template,gene specific primers were designed and advantage 2 polymerase mix was used in PCR,of which using porcine genomic DNA as the template:forward primer was designed according to the acquired consensus region of human and pig FGL2 3′ sequences while reverse primer was designed from human FGL2 3′ end downstream sequence;TA cloning.

以猪正常小肠及心脏组织提取新鲜总RNA，反转录后作为模板，设计基因特异性引物，采用Advantage 2 聚合酶混合物进行PCR扩增；依据猪与人FGL2基因3′端已知同源序列设计PCR上游引物，以人FGL2基因3′末端序列设计下游引物，以猪基因组DNA为模板采用Advantage 2 聚合酶混合物进行PCR反应；PCR载体重组质粒DNA亚克隆扩增。

Anguillarum isolates, while 94.3% and 91.9% nucleotide identical to L. pelagius and Photobacterium damselae respectively. MP analysis showed that ayu-H080701 shared 97.6%-98.8 % amino acid sequence identical to L. anguillarum isolates, while lower than 75.6 % to other bacteria. Phylogenetic analysis showed that ayu-H080701 grouped constantly with L. anguillarum isolates. The biochemical, physiological tests and sequence analysis all strongly supported the identification of the pathogen causing ayu vibriosis in Ninghai country, China, as an isolate of L.

PCR扩增检测表明，细菌16S rRNA 基因通用引物和鳗利斯顿氏菌MP基因特异引物均能扩增到预期大小的特异性条带。ayu-H080701与鳗利斯顿氏菌16S rRNA基因核苷酸序列同源性最高，为99.4%～99.5%，与同属的海弧菌和美人鱼发光杆菌分别为94.3%和91.9%；ayu-H080701与鳗利斯顿氏菌MP氨基酸序列同源性高达97.6%～98.8 %，与其它弧菌科成员则低于75.6 %，系统进化树分析也揭示ayu-H080701与鳗利斯顿氏菌进化相关性最高。

On the other hand, the more important thing is because the urban housing is a kind of heterogeneity products.

另一方面，更重要的是由于城市住房是一种异质性产品。

Climate histogram is the fall that collects place measure calm value, cent serves as cross axle for a few equal interval, the area that the frequency that the value appears according to place is accumulated and becomes will be determined inside each interval, discharge the graph that rise with post, also be called histogram.

气候直方图是将所收集的降水量测定值，分为几个相等的区间作为横轴，并将各区间内所测定值依所出现的次数累积而成的面积，用柱子排起来的图形，也叫做柱状图。