Coat protein gene of 12 isolates of Strawberry vein banding virus(SVBV) was studied by multiple sequence alignment and the primers located in conserved region were designed.The detection protocol for SVBV by reverse t...Coat protein gene of 12 isolates of Strawberry vein banding virus(SVBV) was studied by multiple sequence alignment and the primers located in conserved region were designed.The detection protocol for SVBV by reverse transcriptase polymerase chain reaction(RT-PCR) was developed.The primers of multiplex RT-PCR were selected by primer-primer interactions and the melting temperature.The annealing temperature,the concentration of PCR buffer,the extension temperature,the extension time and the concentration of pri-mers were optimized,respectively.A multiplex RT-PCR assay was made for simultaneous detecting Strawberry mottle virus(SMoV),Strawberry mild yellow edge virus(SMYEV) and SVBV.Both field-grown strawberries and microplants were detected effectively.It was the first report that multiplex RT-PCR was used to assay the efficacy of strawberry viruses elimination.展开更多
Background. Melanoma antigen genes (MAGE)and GAGE genes are encoded by genes t hat are silent in virtually all normal adult tissues but are expressed in tumors from various tissues. These gene products are targets for...Background. Melanoma antigen genes (MAGE)and GAGE genes are encoded by genes t hat are silent in virtually all normal adult tissues but are expressed in tumors from various tissues. These gene products are targets for specific immunotherap y as they are presented by HLA I molecules and recognized by autologous cytotoxi c T-lymphocytes. However, the characteristics of these genes, especially in ute rine cervical cancer are relatively unknown. Purpose. This study evaluated the p revalence of MAGE and GAGE by reverse transcription-poly-merase chain reaction (RT-PCR) with common primers and discusses clinical implications in cervical c arcinoma. Materials and methods. Fresh tissue from 37 cases of primary squamous cell carcinoma and normal cervical mucosa were evaluated for clinicopathologic p arameters including Human Papilloma Virus (HPV)-16,18 infection by PCR, tumor s tage by FIGO classification and lymph node involvement. RT-nested PCR for the M AGE and GAGE genes was performed with common primers and DNA sequencing after su bcloning was used for identification of PCR products of MAGE. Formalin-fixed pa raffin embedded material from the same specimen was analyzed by in situ RT-PCR for MAGE. Results. Expression of MAGE and GAGE was not observed in normal tissue s. Eleven out of 37 cases expressed MAGE mRNA (29.7%): analysis of subtypes ide ntified one case of MAGE-1, two cases of MAGE-4b, six cases of MAGE-3, and tw o unknown subtypes. Thirteen out of 37 cases (35.1%) expressed GAGE mRNA. No si gnificant relationships between expression of these genes and FIGO staging, lymp h node metastasis or HPV infection were found. Conclusion. Expression of MAGE an d GAGE may be involved in the development of uterine cervical carcinoma from int raepithelial neoplasia, although without distinct prognostic significance. MAGE and GAGE genes have the potential to be used as targets for the treatment of ute rine cervical carcinoma.展开更多
目的运用实时荧光定量RT-PCR的方法,检测小RNA分子(small interfering RNAs,siRNA)对乙型肝炎病毒(hepatitis B virus,HBV)复制的抑制作用。方法在HepG2.2.15细胞内导入筛选的三条特异抗HBV的siRNA分子,用实时荧光定量RT-PCR的方法,检...目的运用实时荧光定量RT-PCR的方法,检测小RNA分子(small interfering RNAs,siRNA)对乙型肝炎病毒(hepatitis B virus,HBV)复制的抑制作用。方法在HepG2.2.15细胞内导入筛选的三条特异抗HBV的siRNA分子,用实时荧光定量RT-PCR的方法,检测干扰后HBV的mRNA表达水平。结果导入特异siRNA分子的细胞中HBV的mRNA表达量明显降低。结论实时荧光定量RT-PCR技术能准确可靠的检测mRNA的表达水平,对siRNA分子干扰效果的评价有很好的应用价值。展开更多
文摘Coat protein gene of 12 isolates of Strawberry vein banding virus(SVBV) was studied by multiple sequence alignment and the primers located in conserved region were designed.The detection protocol for SVBV by reverse transcriptase polymerase chain reaction(RT-PCR) was developed.The primers of multiplex RT-PCR were selected by primer-primer interactions and the melting temperature.The annealing temperature,the concentration of PCR buffer,the extension temperature,the extension time and the concentration of pri-mers were optimized,respectively.A multiplex RT-PCR assay was made for simultaneous detecting Strawberry mottle virus(SMoV),Strawberry mild yellow edge virus(SMYEV) and SVBV.Both field-grown strawberries and microplants were detected effectively.It was the first report that multiplex RT-PCR was used to assay the efficacy of strawberry viruses elimination.
文摘Background. Melanoma antigen genes (MAGE)and GAGE genes are encoded by genes t hat are silent in virtually all normal adult tissues but are expressed in tumors from various tissues. These gene products are targets for specific immunotherap y as they are presented by HLA I molecules and recognized by autologous cytotoxi c T-lymphocytes. However, the characteristics of these genes, especially in ute rine cervical cancer are relatively unknown. Purpose. This study evaluated the p revalence of MAGE and GAGE by reverse transcription-poly-merase chain reaction (RT-PCR) with common primers and discusses clinical implications in cervical c arcinoma. Materials and methods. Fresh tissue from 37 cases of primary squamous cell carcinoma and normal cervical mucosa were evaluated for clinicopathologic p arameters including Human Papilloma Virus (HPV)-16,18 infection by PCR, tumor s tage by FIGO classification and lymph node involvement. RT-nested PCR for the M AGE and GAGE genes was performed with common primers and DNA sequencing after su bcloning was used for identification of PCR products of MAGE. Formalin-fixed pa raffin embedded material from the same specimen was analyzed by in situ RT-PCR for MAGE. Results. Expression of MAGE and GAGE was not observed in normal tissue s. Eleven out of 37 cases expressed MAGE mRNA (29.7%): analysis of subtypes ide ntified one case of MAGE-1, two cases of MAGE-4b, six cases of MAGE-3, and tw o unknown subtypes. Thirteen out of 37 cases (35.1%) expressed GAGE mRNA. No si gnificant relationships between expression of these genes and FIGO staging, lymp h node metastasis or HPV infection were found. Conclusion. Expression of MAGE an d GAGE may be involved in the development of uterine cervical carcinoma from int raepithelial neoplasia, although without distinct prognostic significance. MAGE and GAGE genes have the potential to be used as targets for the treatment of ute rine cervical carcinoma.
文摘目的运用实时荧光定量RT-PCR的方法,检测小RNA分子(small interfering RNAs,siRNA)对乙型肝炎病毒(hepatitis B virus,HBV)复制的抑制作用。方法在HepG2.2.15细胞内导入筛选的三条特异抗HBV的siRNA分子,用实时荧光定量RT-PCR的方法,检测干扰后HBV的mRNA表达水平。结果导入特异siRNA分子的细胞中HBV的mRNA表达量明显降低。结论实时荧光定量RT-PCR技术能准确可靠的检测mRNA的表达水平,对siRNA分子干扰效果的评价有很好的应用价值。
