猪链球菌2型粘附相关因子MRP、FBPS和CPS2J荧光定量PCR检测方法的建立.pdfVIP

Agricultural Science Technology, 2013, 14(10): 1378-1382 Copyright 訫 2013, Information Institute of HAAS. All rights reserved Molecular Biology and Tissue Culture A Method for Detecting Adhesive Related鄄Factors of Streptococcus suis Serotype 2 by Real鄄time PCR 鄢 Wei WANG, Kongwang HE , Yanxiu NI, Junming ZHOU, Xuehan ZHANG, Zhengyu YU, Lixin LU, Aihua MAO, Libin WEN, Xiaomin WANG, Bin LI, Rongli GUO Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Key Laboratory of Veterinary Biologicals Engineering and Technology, Ministry of Agriculture, National Center for Engineering Research of Veterinary Bio鄄products, Nanjing 210014, China Abstract [Objective] This study aimed to establish a method for quantitative detection 猪 链 球 菌 2 型 粘 附 相 关 因 子 of mRNA transcriptional level of SS2 adhesive related鄄factors of Streptococcus suis serotype 2 (SS2) by fluorescent quantitative PCR. [Method] The gene fragments en鄄 MRP 、FBPS 和 CPS2J 荧 光定 coding SS2 adhesive related鄄factors MRP, FBPS and CPS2J and a housekeeping 量 PCR 检测方法的建立 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 汪伟 ，何孔旺 * ，倪 艳秀 ，周俊明 ，张雪寒 ，俞正 constructed, and used as templates in Real鄄time PCR. [Result] Dynamic curves, stan鄄 玉 ，吕立新 ，茅爱华 ，温立斌 ，王小敏 ，李彬 ，郭 dard curves and melting curves of the adhesive related鄄factors and aroA were ob鄄 tained by the optimized Real鄄time PCR system. The standard curves showed a good 容莉 （江 苏省农业科学院兽医研究所 ，农业 linear relationship between template copy number and circulation number, and the 部兽用生物制品工程技术重点实验室 ，国家兽 2 correlati