To construct the suicide vector of hhd B gene marker-free mutant in Haemophilus parasuis( HPS),two pairs of specific primers were designed and synthesized according to the hhd B gene upstream and downstream sequences ...To construct the suicide vector of hhd B gene marker-free mutant in Haemophilus parasuis( HPS),two pairs of specific primers were designed and synthesized according to the hhd B gene upstream and downstream sequences of HPS published in Gen Bank. The hhd B gene upstream and downstream sequences were amplified by PCR,which were further ligated( hhd B-up + down) through overlapping PCR method. NotⅠand SalⅠrestriction enzyme sites were introduced on both ends of the ligated sequence. After the corresponding digestion,the hhd B-up + down sequence was directionally cloned to the suicide plasmid vector p EMOC2. Results showed that the suicide vector of hhd B gene marker-free deleted( p EMOC2Δhhd B) with stable inheritance in E. coli β2155 strain was successfully obtained,thereby laying the foundation for construction of HPS-hhd B gene marker-free mutant strain.展开更多
Haemophilus parasuis（HPS） is one of the most important bacterial infectious diseases harming China＇s pig industry for the last few years.To understand the prevalence status of glasser＇s disease in China, the disea...Haemophilus parasuis（HPS） is one of the most important bacterial infectious diseases harming China＇s pig industry for the last few years.To understand the prevalence status of glasser＇s disease in China, the disease samples submitted by Shandong, Henan, Heilongjiang, Jiangxi, Liaon-ing, Guangdong, Shaanxi and Sichuan provinces were isolated and identified. The synovial fluid, lung, brain tissue, heart blood and nasal cavity ofsick pigs in 194 pig farms collected from 2006 to 2009 were detected, and 47 strains of HPS were eventually isolated, with the separation rate of24.2%. Serotypes of the isolates were identified through KRG （Kieletein-Rapp-Gabriedson） agar diffusion method. There were 13 strains of serotype4 （27.7%）, 10 strains of serotype 5 （21.3%）, 8 strains of serotype 12 （17%） and7 strains of serotype 13 （14.9%）, indicating the major serotypes ofHPS in China were serotype 4, serotype 5, serotype 12 and serotype 13.展开更多
为研究来自河南省不同猪源的副猪嗜血杆菌（Hps）的遗传演化关系,应用PCR方法扩增所分离的良种猪源、家养野猪源和地方土猪源等3株Hps（KF0901、JZ0801和XY0501）的16S r RNA基因,并进行序列比较分析。结果 3株Hps的16S r RNA基因全长均...为研究来自河南省不同猪源的副猪嗜血杆菌（Hps）的遗传演化关系,应用PCR方法扩增所分离的良种猪源、家养野猪源和地方土猪源等3株Hps（KF0901、JZ0801和XY0501）的16S r RNA基因,并进行序列比较分析。结果 3株Hps的16S r RNA基因全长均为822 bp,彼此间核苷酸序列同源性为99.3%~99.6%,与参考菌株的核苷酸序列同源性为97.1%~99.4%;基于16S r RNA基因序列绘制的系统进化树显示,本研究中家养野猪源Hps和地方土猪源Hps均属于血清5型,而良种猪源血清5型Hps却与血清12型和血清14型的亲缘关系更近。表明Hps在良种猪、家养野猪和地方土猪之间彼此交叉感染或具有共同来源;并非所有血清5型Hps分离株都属同一分支。展开更多
基金Supported by Science and Technology Project of Guangdong Province(2013B020307002)
文摘To construct the suicide vector of hhd B gene marker-free mutant in Haemophilus parasuis( HPS),two pairs of specific primers were designed and synthesized according to the hhd B gene upstream and downstream sequences of HPS published in Gen Bank. The hhd B gene upstream and downstream sequences were amplified by PCR,which were further ligated( hhd B-up + down) through overlapping PCR method. NotⅠand SalⅠrestriction enzyme sites were introduced on both ends of the ligated sequence. After the corresponding digestion,the hhd B-up + down sequence was directionally cloned to the suicide plasmid vector p EMOC2. Results showed that the suicide vector of hhd B gene marker-free deleted( p EMOC2Δhhd B) with stable inheritance in E. coli β2155 strain was successfully obtained,thereby laying the foundation for construction of HPS-hhd B gene marker-free mutant strain.
文摘Haemophilus parasuis（HPS） is one of the most important bacterial infectious diseases harming China＇s pig industry for the last few years.To understand the prevalence status of glasser＇s disease in China, the disease samples submitted by Shandong, Henan, Heilongjiang, Jiangxi, Liaon-ing, Guangdong, Shaanxi and Sichuan provinces were isolated and identified. The synovial fluid, lung, brain tissue, heart blood and nasal cavity ofsick pigs in 194 pig farms collected from 2006 to 2009 were detected, and 47 strains of HPS were eventually isolated, with the separation rate of24.2%. Serotypes of the isolates were identified through KRG （Kieletein-Rapp-Gabriedson） agar diffusion method. There were 13 strains of serotype4 （27.7%）, 10 strains of serotype 5 （21.3%）, 8 strains of serotype 12 （17%） and7 strains of serotype 13 （14.9%）, indicating the major serotypes ofHPS in China were serotype 4, serotype 5, serotype 12 and serotype 13.
文摘为研究来自河南省不同猪源的副猪嗜血杆菌（Hps）的遗传演化关系,应用PCR方法扩增所分离的良种猪源、家养野猪源和地方土猪源等3株Hps（KF0901、JZ0801和XY0501）的16S r RNA基因,并进行序列比较分析。结果 3株Hps的16S r RNA基因全长均为822 bp,彼此间核苷酸序列同源性为99.3%~99.6%,与参考菌株的核苷酸序列同源性为97.1%~99.4%;基于16S r RNA基因序列绘制的系统进化树显示,本研究中家养野猪源Hps和地方土猪源Hps均属于血清5型,而良种猪源血清5型Hps却与血清12型和血清14型的亲缘关系更近。表明Hps在良种猪、家养野猪和地方土猪之间彼此交叉感染或具有共同来源;并非所有血清5型Hps分离株都属同一分支。
