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1.
小鼠肾素-1和肾素-Ⅱ的cDNA有96%的同源性,以XhoⅠ从重组质粒PKG-R2D上切取包含Ren-2全长基因及周围序列的24kb片段。该片段分别经单、双酶切后,用Ren-1cDNA半长及全长序列为探针进行分子杂交,据片段大小及杂交带的位置关系构建出此24kb片段的物理图谱。  相似文献   

2.
对pDJH2片段DNA进行了序列测定,结果:插入片段为1811个核苷酸;可读框2个:ORF1,1-565bp;ORF2(从最后一个密码倒算)1-662bp,计算机序列同源性或类似性匹配(alignment)表明,ORF1与ORF2的核苷酸类似性为49.36%;ORF1与L.kirschneri(L.alstoni)流感伤寒型omp核苷酸序列类似性为49.26%;ORF2与L.alstoni类似性为51.56%;ORF1与其它细菌(包括螺旋体)omp核苷酸序列对准比较,序列类似性均在43%以上。经查阅EMBLDNA序列数据库未见类似序列。作者认为pDJH2的1.9kb插入DNA片段(ORF1与ORF2)可能是具有保护作用的omp基因片段。  相似文献   

3.
利用CD23cDNA全基因克隆pUCD976,分别以限制性内切酶EcoRI和XbaI酶切该重组质粒DNA,回收1.0kb的CD23cDNA全基因、HindⅢ酶切后回收3’端607bp的结合区段基因以及EcoRⅠ和HindⅡ酶切回收5’端403bp的调控区段基因。将各回收片段先以Klenow酶补平后,插入已补平的pZIP逆转录病毒载体BamHⅠ单切点,利用地高辛素标记的CD23cDNA全基因探针,以斑点核酸杂交筛选逆转录病毒重组体,结合用限制性内切酶BamHI和/或BglⅡ酶切鉴定插入方向,最终获得正、反向插入的CD23全基因-逆转录病毒重组体各2个,反向插入3’端和5’端部分基因片段的重组体分别为4个和1个。为进行CD23反义RNA的研究和真核表达CD23提供了可靠的物质基础。  相似文献   

4.
从人白细胞中分离DNA,经EcoRI酶切后,进行L-mycSouthern杂交,得出人L-myc限制性内切酶片段长度多态性(RFLP)图谱。结果发现,经EcoRI酶切产生L-带(10kb)及S-带(6kb)。对L-mycRFLP与肺癌恶变与预后的关系进行了讨论。  相似文献   

5.
重组丙型肝炎病毒基因载体的构建与表达   总被引:1,自引:1,他引:0  
应用基因重组技术。从载体CDZ2酶切获得1.73kbDNA片段(含16d93bp的HCV结构区cDNA,其特异性已为southernblot证实)。将HCVcUNA片段插入载体pcDNA3,构建HCV重组体。经在大肠杆菌中表达,筛选、酶切分析,获得重组HCV(PcDNA3-HCV)。以重组质粒转染H9细胞(CD淋巴细胞系),用G418筛选出抗性细胞。经RT-PCR、dot-ELISA验证表明,pcDNA3-HCV能在H9细胞中进行复制、转录和表达。这为丙型肝炎发病机理的研究提供了一个有用模型。  相似文献   

6.
目的应用分子遗传学方法对患者在基因水平上作出诊断。方法应用dystropincDNA14kb探针(包括6个亚探针1-2a,2b-3,4-5a,5b-7、8、9-14)与一名18岁的临床表现温和肌营养不良患者及2例对照者(1例正常25岁男性及1例12岁DMD患者)的基因组DNA/HindⅢ片段进行Southern印迹分析。结果前者在与亚探针5b-7杂交中,发现其1.5kb,0.5kb杂交带缺失,在与亚探针8杂交中,发现10.0kb杂交带缺失,表明这三个Hd片段分别含有dystrophin基因的45、46、47号外显子。结论证明患者缺失了45、46、47号外显子,故诊断该患者为Becker型肌营养不良,从而为该家系的遗传咨询获得了可靠的资料。  相似文献   

7.
用DMDcDNA8探针从假肥大型肌营养不良(Duchennemusculardystrophy,DMD)基因的YAC克隆的cosmid亚克隆库中筛选到9个阳性cosmid克隆,Southern杂交鉴定cosmidC0461含DMD基因的第51号外显子,将该cosmid亚克隆于pVC118中,获得含DMD基因第50和51号内含子的3.1kb-HindⅢ片段的亚克隆,将亚克隆的插入片段分别用Sau3AⅡ完全和部分酶切克隆于pUC118中,Sanger法双链测序测出质粒克隆的序列并重叠所测出的序列,确定该片段的长度为3179bp。与Speer测出的该段序列(3159bp)比较相差20bp,有33处不同。并发现有重复顺序存在,它们之间可能会发生重组并可能导致第51号外显子的缺失。  相似文献   

8.
nm_(23)是一种有效的肿瘤转移抑制基因,目前已发现的两种人类nm_(23)(H_1和H_2)的氨基酸肽具有88%的同源性。我们采用PCR技术从人肝基因组DNA中扩增nm_(23)序列,经克隆筛选得到5’端375bp克隆,命名为nm_(23)-H3b。序列分析发现,nm23-H_3b在40bp~70bp之间与原序列完全不同,其他序列与nm_(23)-H1有86%的同源性,与nm_(23)-H2有90%的同源性。BglII消化人肝基因组DNA,用nm_(23)-H_3bDNA为探针,经SouthernBlot后发现10.5、7.9和4.0kb的3条杂交带,未见与nm_(23)-H_1和H_2相同的杂交带。因此认为nm_(23)-H_3b是一种与人肿瘤转移抑制基因nm_(23)-H_1和H_2高度同源的新基因克隆。  相似文献   

9.
根据已知的日本血吸虫菲律宾株的副肌球蛋白分子的部分cDNA序列,设计两对寡核苷酸引物(引物1/2和引物3/4),以聚合酶链式反应(PCR),用引物1/2从本室两个日本血吸虫中国大陆株cDNA库中均扩增出与预期大小(927bp)一致的特定DNA片断。巢式PCR-以第一扩增产物为模板,用引物3/4扩增出约5000bp的单一条带,与预期片段(494bp)大小一致。表明PCR产物为编码副肌球蛋白的目的基因  相似文献   

10.
根据已知的日本血吸虫菲律宾株的副肌球蛋白分子的部分cDNA序列,设计两对寡核苷酸引物(引物1/2和引物3/4),以聚合酶链式反应(PCR),用引物1/2从本室两个日本血吸虫中国大陆株cDNA库中均扩增出与预期大小(927bp)一致的特定DNA片段。巢式PCR——以第一扩增产物为模板,用引物3/4扩增出约500bp的单一条带,与预期片段(494bp)大小一致。表明PCR产物为编码副肌球蛋白的目的基因片段。  相似文献   

11.
Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P < 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (>0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P <0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P < 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.  相似文献   

12.
Objective: To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods: SD rats served as both donors and recipients. 4℃ sodium lactate Ringer's was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava (IVC) inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results: Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion: This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.  相似文献   

13.
FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…  相似文献   

14.
Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P< 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P< 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.  相似文献   

15.
Shock wave lithotripsy (SWL) is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it's difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.  相似文献   

16.
Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.  相似文献   

17.
INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5  相似文献   

18.
People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.  相似文献   

19.
Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.  相似文献   

20.
In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.  相似文献   

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