[Objective] This study aimed to establish a method for quantitative detection of mRNA transcriptional level of SS2 adhesive related-factors of Streptococcus suis serotype 2 (SS2) by fluorescent quantitative PCR. []V...[Objective] This study aimed to establish a method for quantitative detection of mRNA transcriptional level of SS2 adhesive related-factors of Streptococcus suis serotype 2 (SS2) by fluorescent quantitative PCR. []Vlethod] The gene fragments en- coding SS2 adhesive related-factors MRP, FBPS and CPS2J and a housekeeping gene aroA were amplified by reverse transcription PCR from the total RNA of SS2, cloned, and sequenced. The recombinant plasmids containing the target genes were constructed, and used as templates in Real-time PCR. [Result] Dynamic curves, stan- dard curves and melting curves of the adhesive related-factors and aroA were ob- tained by the optimized Real-time PCR system. The standard curves showed a good linear relationship between template copy number and circulation number, and the correlation coefficients (FF) of the standard curves were over 0.995. Also, these as- says were highly specific a^d there was single specific melting peak for every gene. Moreover, the assays were highly sensitive and had a detection limit of 1.0×102 copies in 1 μl of initial templates. Finally, it was highly repeatable and had a coeffi- cient of variation less than 2% for intra-assay. [Conclusion] This study will provide a way to reveal the adhesion mechanism of SS2 to different host cells at molecular level.展开更多
随着3G时代的逐步到来,集办公、娱乐、通讯等多功能于一体的智能化手机已成为各移动设备开发商的开发热点。J2ME(Java 2 Mobile Edition)平台被认为是最杰出的手机程序开发平台,J2ME应用程序可运行在不同制造商的不同设备上,真正体现了...随着3G时代的逐步到来,集办公、娱乐、通讯等多功能于一体的智能化手机已成为各移动设备开发商的开发热点。J2ME(Java 2 Mobile Edition)平台被认为是最杰出的手机程序开发平台,J2ME应用程序可运行在不同制造商的不同设备上,真正体现了Sun公司所倡导的"write once,run anywhere"。讨论了在Windows下的J2ME开发环境下的手机(或PDA等手持移动设备)实时监视系统问题,设计并实现了对远程计算机的实时监视。展开更多
基金Supported by National Natural Science Foundation of China(31072155)Natural Science Foundation of Jiangsu Province(BK2010068)+1 种基金Fund for Independent Innovation of Agricultural Science in Jiangsu Province[CX(11)2060]Special Fund for Agroscientific Research in the Public Interest(201303041)~~
文摘[Objective] This study aimed to establish a method for quantitative detection of mRNA transcriptional level of SS2 adhesive related-factors of Streptococcus suis serotype 2 (SS2) by fluorescent quantitative PCR. []Vlethod] The gene fragments en- coding SS2 adhesive related-factors MRP, FBPS and CPS2J and a housekeeping gene aroA were amplified by reverse transcription PCR from the total RNA of SS2, cloned, and sequenced. The recombinant plasmids containing the target genes were constructed, and used as templates in Real-time PCR. [Result] Dynamic curves, stan- dard curves and melting curves of the adhesive related-factors and aroA were ob- tained by the optimized Real-time PCR system. The standard curves showed a good linear relationship between template copy number and circulation number, and the correlation coefficients (FF) of the standard curves were over 0.995. Also, these as- says were highly specific a^d there was single specific melting peak for every gene. Moreover, the assays were highly sensitive and had a detection limit of 1.0×102 copies in 1 μl of initial templates. Finally, it was highly repeatable and had a coeffi- cient of variation less than 2% for intra-assay. [Conclusion] This study will provide a way to reveal the adhesion mechanism of SS2 to different host cells at molecular level.