序列性

This result showed that the nucleotide sequence of the coding region and accordingly amino acid sequences of Myf-5 gene of gayal and cow, chimpanzee, rhesus monkey, human, dog, house mouse, Chinese softshell, chicken and zebrafish were quite conservative.

大额牛与普通牛、黑猩猩、恒河猴、人、狗、家鼠、软体贝壳、鸡、斑马鱼的Myf-5基因编码区核苷酸序列同源性分别为99.0％、90.0％、90.5％、90.5％、87.2％、84.8％、72.9％、64.3％、51.8％，相应的氨基酸序列同源性分别为99.6％、94.9％、94.6％、94.9％、89.9％、88.7％、81.6％、70.4％、56.6％，这说明大额牛与其他9个物种在Myf-5基因的编码区核苷酸和相应氨基酸序列上具有较高的保守性。

The cattle BMPRⅠA gene full-long cDNA is 4067bp, which encodes 532 amino acids and shares 91%, 93%, 89%, 88%, 91% and 82% similarity in nucleic acid sequences and 97%, 95%, 96%, 95%, 97% and 91% similarity in amino acid sequences with human, chimpanzee, mouse, rat, dog and red jungle fowl, respectively.

克隆得到了牛BMPRⅠA基因4067bp的cDNA全长序列，通过核酸序列分析发现，该基因编码532个氨基酸，与人、黑猩猩、小鼠、大鼠、犬和红原鸡在核酸序列上分别有91%，93%，89%，88%，91%和82%的同源性；在氨基酸序列上分别有97%，95%，96%，95%，97%和91%的同源性。

A clone from each genotype was randomly selected as representative for sequencing. The obtained 16S rDNA gene sequences had a similarity of 87%-100% with those in the GenBank (www. ncbi. nlm. nih. gov), and more than half of them had a similarity lower than 97%, being of new species. Based on phylogenetic analysis, the bacteria in the two soils were classified into 10 groups, with 5 groups in common. The dominant bacterial groups in the two soils differed obviously. In primeval forest soil, the dominant group was Proteobacteria, which had 39 genotypes, occupying 58.0% of all the clones; while in the soil of degraded ecosystem the dominant groups were Acidobacteria and Proteobacteria, which had 19 and 15 genotypes occupying 32.5% and 30.5% of all the clones, respectively. In the soil of degraded ecosystem, Proteobacteria group decreased while Acidobacteria group increased markedly, compared with those in primeval forest soil.

从每种基因型中随机选择一个克隆子作为代表进行测序分析，所有序列与GenBank数据库中序列的同源性为87%~100%，且两样地中均有超过一半的基因型序列与数据库中已知序列同源性低于97%，属于分类在"种"地位上的新发现细菌；通过系统发育研究将两样地的细菌分为10大类群，两样地共同拥有5大类群，但两样地的细菌优势类群明显不同，原生土壤为Proteobacteria，含39种基因型，占总克隆子数的58.0%，退化生态系统土壤为Acidobacteria和Proteobacteria，分别含19种和15种基因型，占总克隆子数的32.5%和30.5%；与原生土壤细菌类群相比，退化生态系统土壤Proteobacteria类群明显减少，Acidobacteria类群明显增加。

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。

A model for finding the cryptic period of hydrological time series is proposed. The functions of the model include to apply logarithm transformation for making the sequence more stationary, to construct a corresponding auto-regression model for reducing the herteroskedasticity of residual, to determine the component of the prime period by periodogram analysis.

本文在讨论水文时序平稳性和异方差性的基础上，提出了检测水文时序隐含周期的模型，即对水文时序设计对数变换序列，提高序列的平稳性；建立相应的自回归模型降低残差的异方差性；通过对所得平稳序列进行周期图分析确定可能的主周期分量。

Model for finding the cryptic period of hydrological time series is proposed. The functions of the model include to apply logarithm transformation for making the sequence more stationary, to construct a corresponding auto-regression model for reducing the herteroskedasticity of residual, to determine the component of the prime period by periodogram analysis.

bstract 本文在讨论水文时序平稳性和异方差性的基础上，提出了检测水文时序隐含周期的模型，即对水文时序设计对数变换序列，提高序列的平稳性；建立相应的自回归模型降低残差的异方差性；通过对所得平稳序列进行周期图分析确定可能的主周期分量。

The results showed that Me-c84 shared 98% similarities with the 26S ribosomal RNA gene from Sarracenia purpurea, while M14-86 shared97% similarities with the 26S ribosomal RNA gene from Portulaca Grandiflora.

将已经获得的两条cDNA序列与NCBI网站上的GenBank数据库进行同源序列比对，结果显示Me-c84核苷酸序列与瓶子草等植物的26S核糖体RNA部分基因同源性达到98％。M14-86核苷酸序列与大花马齿苋等植物的26S核糖体RNA部分基因同源性达到97％。

OPY13-661 sequence has identity with 16 EST sequences from European grapes, and shows 89% identity with 2 EST sequences from abiotic stressed leaves of Vitis vinifera var. Chardonnay.

OPY13-661序列与欧洲葡萄16条EST序列有同源性，其中与"赤霞珠"cDNA克隆的EST有较高的同源性，与"霞多丽"的2条叶片非生物胁迫有关的EST序列同源性均为89%。

Furthermore, out of 497 fAFLP markers, 80 special bands were found to be able to distinguish the four groups from each other and may be applied for germplasm characterization and molecular assistant classification of Meretrix clam.4 Molecular classification of two species of Meretrix clam based on fAFLP and ITS sequences4.1 The results of fAFLP maker analysis of S, G and W showed that each group had their own specific loci among which there were 53 special loci in W group, much more than those of S group (14) and G group (21). Among the 53 loci, nine were all dominant loci. These unique loci could be taken as molecular markers to distinguish W from other groups. The genetic similarity indexes and distance matrix between S and G groups were 0.9585 and 0.0424 respectively, but the genetic similarity indexes and distance matrix between W group and S or G group was 0.7939 or 0.7941, and 0.2308 or 0.2305 respectively. The results revealed that significant difference existed between W and S or G groups in molecular genetic structure. The phylogenetic trees by the methods of UPGMA and NJ also indicated that S and G populations were very closely related, while W population was a relatively independent cluster, lying beyond the species which S and G belong to.4.2 The internal transcribed spacer region of the rDNA from S group, G group and W group were PCR amplified and sequenced. The results showed that the size of ITS ranged between 1266-1269bp in W group, while those in G and S groups were 1614bp and 1520bp respectively. The GC content ranged 62.32-62.62% in W group while it was 61.77% in G group. The genetic distances between three populations of W group were 0.001～0.003, but it was 0.110 or 0.147 respectively between W group and G group or S group. Phylogenetic trees by NJ method also showed that G group was very closely related to S group, while W group was a relatively independent cluster.

在457个总扩增位点中找出了53个W的特有位点，远多于S群体(14)和G(21)群体，而且在53个特有位点中有9个出现频率为100%的位点，这些位点可以作为区分其它2个群体的特征性标记；S– G群体特有的位点有112个，其中有4个位点出现频率为100%，可作为S– G群体区别于W群体的特征性标记。S群体和G群体间的遗传相似性系数为0.9585，遗传距离只有0.0424，在NJ和UPGMA法构建的亲缘关系的树状图上均首先聚在一起，说明二者的亲缘关系很近，应属于种内群体间的关系；而W与S和G的遗传相似性系数均较小(0.7939和0.7941)，相对遗传距离很大而且十分相近(0.2308和0.2305)，在亲缘关系树状图上单独分出一支，也表明W与S和G群体间的亲缘关系较远。4.2 ITS序列比较分析通过对白壳文蛤、山东文蛤和广西文蛤的ITS序列扩增电泳、PCR-RFLP分析和ITS序列分析发现，W的ITS序列长度在1266-1269 bp，而S和与G的ITS序列总长度分别为1520 bp和1614 bp；从ITS1和ITS2长度来看，W分别为739-741 bp和316-317 bp，S为895 bp和414 bp，G为987 bp和416 bp；而从ITS碱基组成来看，W的GC含量在62.32-62.62%之间，而G群体为61.77%。W的3个壳色不同群体间的遗传距离仅0.001、0.002和0.003，S与G群体间的遗传距离是0.010，说明W群体内变异很小，而S与G群体间已出现明显的遗传分化，但还均属于种内群体间的遗传变异；而W与G和S的遗传距离分别达到0.110、0.147，两个类群差异显著，已远超出种内群体间的遗传变异。

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。