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find Keyword "序列测定" 2 results
  • Sequence Determination and Bioinformatics Analysis of Hp1501 Gene from Helicobacter Pylori NCTC11637

    目的 幽门螺杆菌NCTC11637菌株Hp1501基因进行序列测定,并运用生物信息学软件对其进行分析。 方法 用聚合酶链反应方法从幽门螺杆菌 NCTC11637菌株基因组DNA扩增Hp1501基因,T-A克隆,鉴定后测序,将基因序列向GeneBank提交并申请登录号,用生物信息学软件分析其生物学特性。 结果 成功获取幽门螺杆菌 NCTC11637株Hp1501基因序列,获得GeneBank登录号JF820815。软件分析表明,序列全长为1 164 bp,与幽门螺杆菌国际标准株26695和J99的基因序列一致性为96%~97%,氨基酸序列一致性为97%~98%;软件预测其编码的前59个氨基酸为信号肽,其编码的核心肽为幽门螺杆菌外膜蛋白。 结论 成功测定幽门螺杆菌 NCTC11637菌株Hp1501基因序列并预测出其生物学特性,为进一步研究其功能,阐明其致病机制奠定了基础。Objective To determine the sequence of Hp1501 gene from H. pylori NCTC11637 and analyze the sequence with bioinformatics software. Methods Polymerase chain reaction (PCR) was used to amplify the Hp1501 gene from chromosomal DNA of H. pylori NCTC11637. After T-A clone, the amplified DNA sequence was determined and the gene sequence was sent to GeneBank for analysis with bioinformatics software. Results Hp1501 gene from H. pylori NCTC11637 was successfully attained, and the logging number for GeneBank was JF820815. The analysis showed that the gene sequence was 1164 bp in length, 96%-97% identical in DNA sequence and 97%-98% identical in amino acid sequence compared with standard strain 26695 and J99. The forward 59 amino acids were signal peptide and the core peptide was outer membrane protein of H. pylori based on the software prediction. Conclusions The sequence and bionomics of Hp1501 gene from H. pylori NCTC11637 has been determined successfully. It provides a solid base for the further research of the biological function and pathogenicity mechanism of H. pylori.

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  • 风湿性二尖瓣狭窄心房颤动患者HCN4基因cDNA序列测定及mRNA的表达

    摘要: 目的 通过分析风湿性心脏病二尖瓣狭窄患者心房肌组织超级化激活环核苷酸调控通道基因家族4(HCN4)基因表达与心房颤动发生的关系,为探讨心房颤动发生的机制奠定理论基础。 方法 52例风湿性二尖瓣狭窄患者,根据是否合并心房颤动将其分为两组,实验组:38例,男18例,女20例;年龄26~68岁,平均年龄46.47岁;均合并心房颤动。对照组:14例,男6例,女8例;年龄21~62岁,平均年龄42.93岁;不合并心房颤动。提取并逆转录两组患者心房肌组织中HCN4基因的总核糖核酸(RNA),应用SYBR GreenⅠ荧光染料, 建立检测 HCN4基因信使RNA(mRNA)的实时荧光定量聚合酶链反应 (PCR)法,并对PCR产物测序进行分析。根据标准曲线计算出两组心房肌组织中HCN4基因 mRNA含量,并以HCN4基因mRNA和内参β肌动蛋白(β-actin)含量的比值作为评价HCN4基因mRNA表达水平指标。 结果 测定HCN4基因cDNA 序列同源性为100%。建立的实时荧光定量 PCR方法在103~107拷贝数/μl的标准品梯度稀释范围内r为0.999。实验组HCN4基因mRNA与β-actin含量的相对表达值比值与对照组比较明显升高(1.323±1.226 vs. 0.116±0.192,P<0.05)。 结论 实时荧光定量PCR对HCN4基因mRNA能进行准确定量,HCN4基因的过度转录表达提示其可能参与了调控风湿性二尖瓣狭窄心房颤动的发生过程。

    Release date:2016-08-30 06:01 Export PDF Favorites Scan
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