核苷酸酶

The results showed that:(1) 5'-nucleotide contents in yeast autolysate was slightly increased by adding RNase frommalted barley roots;(2) higher pH did not improve the 5'-nucleotide contents even in the presence of RNase from malted barleyroots, but at the pH 8.0, the addition of Acalase could increase the final yield, soluble protein contents and 5'-nucleotide inthe presence of RNase from malted barley roots.

酵母核酸酶的pH稳定性和温度稳定性均比麦芽根核酸酶的要差。这说明自溶法生产酵母提取物时，即使外加5'-磷酸二酯酶不能有效提高呈味核苷酸的原因并非由于外加的5'-磷酸二酯酶性质不稳定，而是另有其它原因。

Our results showed that 1, 8-Naphthyl imide could improve the properties of natural oligonucleotide in resisting nuclease and binding the targeted sequence by covalently linkage at the end of oligonucleotide. Moreover, our results also showed that the antigene oligonucleotide could specifically cleave the targeted double DNA sequence by modification of naphthyl imide at the 3'end of oligonucleotide.

体外结果表明萘二酰亚胺对寡核苷酸的共价修饰可以提高天然寡核苷酸抗核酸酶降解能力，提高寡核苷酸与靶序列结合的稳定性以及使寡核苷酸具有选择性结合和切断靶DNA的功能。

Antifolate Alimta derived from pyrrolo[2,3-d]pyrimidine is a multitargeted antitumor agent, it can inhibit at least five major folate-dependent enzymes: thymidylate synthase, dihydrofolate reductase, glycinamide ribonucleo -tide formyltransferase, aminoimidazole ribonucleotide formyl transferase and C-1 tetrahydrofolate synthetase(C1-S).

吡咯并[2，3-d]嘧啶类叶酸拮抗剂Alimta是一种多靶向的抗癌药物，它能抑制至少五种叶酸依赖性酶：胸苷合成酶、二氢叶酸还原酶、叶酰多聚谷氨酸合成酶、甘氨酰胺核糖核苷酸转甲酰酶及C1-四氢叶酸合成酶(C1-S)。。

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 cDNA document of ACC oxidase gene of Citrus aurantium L. was cloned. By using the program of BLAST on NCBI GenBank database, the sequence presented a very high match with the ACC oxidase genes from other plants. The base sequence was analysed by using biology programe of DNAStar 5.0. 278 amino acids were coded by the base sequence and the base sequence had the same conserve region of ACC oxidase gene of many kinds of other plants. The base sequences comparability was more than 72% compared with those of many kinds of other plants. The amino acid sequences comparability was more than 70% compared with those of a lot of other plants.

克隆了酸橙1-氨基环丙烷－1-羟酸氧化酶基因cDNA片段，将片段序列在NCBI网站上进行同源性搜索，显示的皆为不同植物的ACC氧化酶基因，因而认为所克隆的片段就是酸橙ACC氧化酶基因；并运用DNAStar5.0软件进行序列分析，推导的氨基酸序列为278个残基；具有所有植物ACC氧化酶基因共有的保守区域；与多种植物的ACC氧化酶基因的核苷酸和氨基酸序列的同源性都在72%和70%以上。

Methods: Five 10-23 deoxyribozymes (DZ1-DZ5) with 2 phosphorothioate groups at 5' and 3' end were designed and synthesized. Full sequence ET-1 RNA was transcripted and FAM-labeled 10-23 deoxyribozyme was used to select cleavable 10-23 deoxyribozyme in vitro and to detect intracellular uptake. The content of ET-1 mRNA was measured by semi-quantitative RT-PCR after 10-23 deoxyribozyme was transfected into cultured neonatal rat cardiomyocytes.

体外转录ET-1全长RNA底物，设计并合成5条ET-1 10-23脱氧核酶(DZ1~DZ5)，其5'及3'端各有2个核苷酸硫代修饰，体外切割ET-1RNA底物，筛选有效脱氧核酶；5'标记荧光素FAM的10-23脱氧核酶瞬间转染新生大鼠心肌细胞以观察对10-23脱氧核酶的摄取；采用半定量RT-PCR检测ET-1基因的表达。

Result 1 Magnetic nanoparticles, magnetic nanoparticles modified with antisense oligodeoxynucleotide of human telomerase reverse transcriptase induced HL-60 tumor cells to apoptosis, we could see typical morphologic change of apoptosis cells: karyopyknosis, chromation"s condensing and aggregation in nuclear, forming crescent-shaped or annulus structures to lean on edge of cell nucleus"s membrane and posing apoptosis body by Atomic Force Microscope, Fluorescence microscope, transmission electron Microscope 2 There was a significant difference compared with control group(p.01), inhibition ratio had significant positive correlation with medication dosage and time ;during 0.8-8μM dosage amplitude, inhibition ratio accrescenced by dosages increasing. However, the inhibition ratio would decrease when dosage over 8-80μM.

结果 1 磁性纳米粒子、修饰有端粒酶反义寡核苷酸的磁性纳米粒子诱导HL-60细胞发生凋亡，原子力显微镜、光学显微镜、荧光显微镜和透射电镜下均观察到HL-60细胞呈现典型的凋亡细胞的形态变化：细胞核固缩，核内染色质浓缩、凝聚、形成新月形或环状结构紧靠在细胞核膜边缘，并形成凋亡小体。2 磁性纳米粒子、修饰有端粒酶反义寡聚脱氧核苷酸的磁性纳米粒子对HL-60肿瘤细胞的生长和增殖有明显的抑制作用，与对照组相比有显著性差异(p＜0.01)，在剂量为0.8-8μmol／L范围内，抑制率随剂量的增加而增加，当剂量超过8μmol／L时，抑制率反而下降；3 磁性纳米粒子、修饰有端粒酶反义寡聚脱氧核苷酸的磁性纳米粒子可增强p53基因的表达活性，引起DNA降解损伤，反向调节细胞周期活动，促使细胞从G0期进入G1期，抑制肿瘤细胞的生长。4 修饰有端粒酶反义寡聚脱氧核苷酸的量子点能通过内吞作用进入HL-60肿瘤细胞的细胞核，可以在细胞内进行定位和促进HL-60肿瘤细胞的凋亡。

This method conformed the following conditions: spleen phosphodiesterase and micrococcal nulcease for digesting reagent and the concentration, temperature and digesting time of the two enzymes;the components of p32-post label alleviant and labeling time;the concentration of p1 nuclease; the concentration of T4 polynucleotide kinase for catalysing and labeling reagent;the components of bidirectional spreader and the spreading time ,etc.

确定了采用脾磷酸二酯酶和微球菌核酸酶消化底物以及两种酶的使用浓度、温度及消化时间；确定了P32标记缓冲液的主要成分及标记时间；确定了P1核酸酶的浓度；确定了T4多核苷酸激酶催化标记底物的浓度；确定了双向展开剂的主要成分及展开时间等。

Micrococcal nuclease hypersensitive DNA fragments were either cloned directly, or mapped onto the genome by hybridization with oligo-nucleotide probes on a 244K Agilent array-CGH chip. The array-CGH mapping was confirmed for nuclease sensitivity by PCR after MNase digestion and by Southern blot analysis.

我们利用小球菌核酸酶超敏感的去氧核醣核酸片段直接选殖或是将其作为寡核苷酸探针於安捷伦244k寡核苷酸阵列-比较基因体杂合晶片上，进行基因体定位，并利用聚合酶链索反应和南方墨点法侦测其核酸酶敏感程度，以确认结果之可靠性。

Methods The DNA chip was fabricated,on which oligonucleotide bougies′length was 19 base pairs,and the concentration was above 15 μmog/L.

方法自制DNA芯片，每条寡核苷酸探针片段长度为19bp，点样浓度在15μmol/L，以痰液为待检标本，分别用不同引物、Mg2+、核苷酸、DNA聚合酶浓度等进行多重聚合酶链反应；分别在不同时间、温度下进行杂交，分别在不同酶作用时间和显色时间进行分析，比较不同条件芯片检测结果。

However, as the name（read－only memory）implies, CD disks cannot be written onorchanged in any way.

然而，正如其名字所指出的那样，CD盘不能写，也不能用任何方式改变其内容。

Galvanizes steel pallet is mainly export which suits standard packing of European Union, the North America. galvanizes steel pallet is suitable to heavy rack. Pallet surface can design plate type, corrugated and the gap form, satisfies the different requirements.

镀锌钢托盘多用于出口，替代木托盘，免薰蒸，符合欧盟、北美各国对出口货物包装材料的法令要求；喷涂钢托盘适用于重载上货架之用，托盘表面根据需要制作成平板状、波纹状及间隔形式，满足不同的使用要求。