内切

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。

Polymarase Chain Reaction may be the better method of detecting M.tuberculosis than other methods and lead us to carry out a series of research on this problem. This thesis is composed of five parts:(1)systematic evaluation for the first time of the ability of five defferent PCR methods to detect M. tuberculosis;(2) using the endonuclease and southern blot by a radioactive and a nonradioactive labeled probe, to confirm the specificity of the PCR products and to analyse the sensitivity of detection by southern blot method using digoxigenin-labeled probe;(3) evaluation of acid-fast staining, bacterial culture and PCR for the detection of M. tuberculosis in sputum, pleural and peritoneal effusion samples;(4) The feasibility of PCR for detection of M tuberculosis on formalin-fixed paraffin embedded tissues;(5) 26 cases of sarcoidosis were retrospectively studiedwith PCR for M. tuberculosis and nontuberculosis Mycobacterium and the problems on histopatholoical diagnosis of sarcoidosis are discussed.

论文共分为五个部分，(1)首次系统地比较了五种不同引物序列的PCR检测结核杆菌方法的特异性和敏感性等情况；(2)利用限制性内切酶酶切分析和同位素及非同位素标记DNA探针进行Southern杂交的办法，分析PCR产物的序列与原设计是否相符及杂交检测PCR产物对提高检测结核杆菌敏感性的情况；(3)并对PCR用于快速检测临床标本中的结核杆菌进行了初步评价；(4)探讨了PCR用于检测石蜡包埋组织中结核杆菌的作用；(5)并利用PCR检测结核杆菌和非结核分枝杆菌的方法对26例原病理诊断为结节病的病例进行了PCR检测及组织病理学诊断的探讨。

Methods:The whole mature protein coding sequence of three truncated dystrophin gene was amplified by RT-PCR method applied to human muscle cDNA. The fragment was inserted into prokaryotic expression vector pET28a plasmid.

以人肌肉组织cDNA为模板，采用RT-PCR扩增三个截短的肌营养不良蛋白cDNA，分别克隆入原核表达载体pET28a中，经限制性内切酶双酶切及DNA序列分析鉴定目的基因。

A human transforming growth 〓 cDNA was inserte in reverse orientation into the retroviral vector, pLXSN. Restriction en zyme analysis was used to confirm the antisense orientation of 〓〓 in the pLXSN vector from individual transfomants.

将人的转化生长因子-〓cDNA（1467bp）反向插入逆转录病毒载体pLXSN，筛选转化菌，挑取抗性克隆，用限制性内切酶酶切鉴定重组子。

After thecomplete genome extraction of the strain was performed, the genomic DNA was partiallydigested by restriction enzyme Sau3AⅠ, the DNA fragments from 1 to 5Kb was clonedinto prokaryote expression vector pET-28a-c, and transformed host bacteria. The resultsshowed that we succeeded in constructing the gene expression library of haemophilusparasuis serovar 5, which is fundamental for the study of advanced gene screening. Inaddition, primer design was performed based on haemophilus influenzae in this study. In addition, PCR was performed by using genomic DNA of haemophilus parasuisserovar 5 as the template. The results demonstrated that we obtained two neo-gene:23SrRNA gene(conserved gene belonging to the large-subunit of ribosome) and adenylatecyclase gene(encodes adenylate cyclase and participates in converting adenyl nucleosidetriphosphate to cyclic adenosine3",5"-monophosphate). Furthermore, the phylogeneticanalyses between the species was performed, and neighbor-joining tree was constructedbased on comparison of 23S rRNA gene sequences, so it was illuminated betweenHaemophilus parasuis and other species in molecular evolution relationship.

选择我国流行优势菌株副猪嗜血杆菌血清5型地方株为研究对象，提取细菌基因组DNA，用限制性内切酶Sau3AⅠ对基因组DNA进行部分酶切，回收大小为1～5Kb的DNA片段，将其连接入原核表达载体pET-28a-c，最后转化宿主菌，结果成功地构建了基因表达文库，为后续的基因筛选工作奠定基础；另外，本研究选择嗜血杆菌属的流感嗜血杆菌为参考对象进行引物的设计，以副猪嗜血杆菌血清5型地方菌株的基因组DNA为模板，进行PCR扩增反应，结果表明成功地获得两个新基因：23S rRNA基因(存在于核糖体大亚基中的保守性基因)和腺苷酸环化酶基因(负责将腺嘌呤核苷三磷酸转变为环腺苷酸)，并进一步做了不同物种之间的分子系统发育分析，构建了基于23S rRNA基因的邻接法系统发育树，阐明了副猪嗜血杆菌与其它菌种的分子进化关系。

Methods With polymerase chain reaction-single-strand conformation polymorphism combining argentation and glue retrieval, DNA sequencing, and restriction fragment length polymorphism, the SNP of the 50 exons in all the coding regions of ABCA1 gene was detected in 112 patients with established diagnosis of coronary heart disease.

通过聚合酶链反应－单链构象多态性分析结合银染后胶回收、DNA测序和限制性内切酶酶切分析方法检测112例明确冠心病诊断患者的ABCA1基因全部编码区50个外显子的SNP。

Methods Trizol reagent from clamworm digestive tract epithelial cells extracted methods Total RNA. The mRNA was purified and reverse transcripted into cDNA. The cDNA was linked into plasmid pDNR-Lib after digested by nucleate endonuclease Sfi I to construct cDNA library.

分离沙蚕消化道上皮细胞，使用Trizol试剂提取总RNA，纯化mRNA，用MMLV逆转录成cDNA，用Sfi I核酸内切酶酶切后连接到pDNR-Lib质粒中，构建cDNA文库。

Any of several enzymes, such as endonucleases and exonucleases, that hydrolize nucleic acids .

核酸酶水解核酸的酶，如核酸内切酶和核酸外切酶

Any of several enzymes, such as endonucleases and exonucleases, that hydrolize ''.

核酸酶水解核酸的酶，如核酸内切酶和核酸外切酶

Any of several enzymes,such as endonucleases and exonucleases,that hydrolize nucleic acids.

核酸□水解核酸的□，如核酸内切□和核酸外切

But we don't care about Battlegrounds.

但我们并不在乎沙场中的显露。

Ah! don't mention it, the butcher's shop is a horror.

啊！不用提了。提到肉，真是糟透了。