检索结果-南通市图书馆

Establishment and Application of a Real-time fluorescent quantitative pcr Method for Detection of Porcine Circovirus Type 2

引用

Animal Husbandry and Feed Science 2015年第4期7卷 249-252,256页

作者： Dong Lin Wei Feng Guan Yu Liu Zengshan Shen Zhiqiang College of Animal Science and Veterinary Medicine Jilin University Shandong Binzhou Animal Science and Veterinary Medicine Academy

[ Objective ] To establish a real-time fluorescent quantitative polymerase chain reaction （pcr） method with SYBR Green I for the detection of porcine circovirus type 2 （PCV2）. [Methods] Specific primers were designed to amplify the conserved gene segments of PCV2 with a size of 177 bp by pcr. The ampli- fied gene was cloned into the vector of pMD 18-T and transformed into DHSct to screen positive clones. After being extracted and purified, the recombinant plasraids pMD 18-T-177 were taken as the standard DNA templates to establish the fluorescence quantitative pcr method for the detection of PCV2, and the pcr re- action conditions were optimized. [ Results] Ct value of the established pcr method showed a good linear relationship with the standard DNA templates within a viral load of 3.21 × 100 -4.16 × 108 copies/μL , the correlation coefficient was O. 998 8 and the slope was - 3.286. The method did not show any cress-reactions with the genomes of PRRSV, PCV1, CSFV, PRV, PPV and Escherichia coli. Sensitivity of this method was proved to be 3.21 × 10 copies/μL, which was 1 000 times higher as conventional pcr method. Variation coefficients of the repeated trims among same batch or different batches were both less than 3.00%. Positive rate of clinical samples detected by the established pcr method was 58.94%, which was significantly higher than the detection rate by conventional pcr. [ Conclusions ] A reM-time fluorescent quantitative pcr method with SYBR Green I for the detection of PCV2 was established, which was better for conducting the quan- titative analysis and the early diagnosis of PCV2 infection.

关键词： Porcine circovirus type 2 fluorescent quantitative pcr SYBR Green I

来源：

维普期刊数据库评论

在线全文

维普期刊数据库

学校读者我要写书评

暂无评论

Development of a real-time pcr method for Thalassiosira rotula rapid detection

引用

Acta Oceanologica Sinica 2007年第2期26卷 133-139页

作者： HE Shanying YU Zhigang MI Tiezhu College of Chemistry and Chemical Engineering Ocean University of China Qingdao 266003 China College of Environmental Science and Engineering Ocean University of China Qingdao 266003 China

Gene specific primers and DNA probe were designed based on the sequence of 18S rDNA cloned from the red tide alga Thalassiosira rotula. A real-time fluorescent quantitative pcr （RFQ - pcr） method was developed for quantitative detection of T. rotula. The RFQ - pcr assay data showed that the results obtained with the RFQ - pcr quite good agreement with those with the light microscope （LM） counting method, which suggested that the RFQ - pcr could be a useful method for red tide alga detection.

关键词： red tide Thalassiosira rotula fluorescent quantitative pcr 18S rDNA

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Estimation of growth rate of Skeletonema costatum based on relative expression amount of PCNA gene

引用

Acta Oceanologica Sinica 2008年第6期27卷 149-158页

作者： HE Shanying YU Zhigang College of Environmental Science and Engineering Zhejiang Gongshang University Hangzhou 310012 China College of Chemistry and Chemical Engineering Ocean University of China Qingdao 266003 China

Partial sequence of Skeletonema costatum proliferating cell nuclear antigen （PCNA） gene was obtained by reverse transcriptase pcr（RT-pcr） and 3＇ rapid amplification of cDNA ends （3＇-RACE） techniques. Based on the obtained PCNA and cytochrome b gene（ Cyt b gene） sequences, a real-time fluorescent quantitative pcr （FQ-pcr） method was developed to detect the expression S. costatum PCNA gene, and this method was applied to study the relationship between the growth rate of S. costatum and the average expression amount of PCNA gene in a single cell. The expression amount of PCNA gene had large variation in cells collected at different culture phases, and the trend was well consistent with the growth rate, which suggested that the expression amount of PCNA gene correlated well with the cell division, and the PCNA could be a promising indicator for the S. costatum cell proliferation. Furthermore, using the PCNA gene as the objective gene and the Cyt b gene as the house-keeping gene, a new method for estimating the in situ growth rate of S. costatum was established by analysis of the relative expression quantity （REQ） of the PCNA gene.

关键词： Skeletonema costatum fluorescent quantitative pcr PCNA growth rate

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Development of real-time pcr method for rapid detection and quantification of Heterosigma akashiwo

引用

Journal of Harbin Institute of Technology(New Series) 2008年第1期15卷 118-123页

作者：何闪英于志刚米铁柱 College of Environmental Science and Engineering Zhejiang Gongshang University College of Chemistry and Chemical Engineering Ocean University of China College of Environmental Science and Engineering Ocean University of China

To rapidly detect the harmful algae *** qualitatively and quantitatively, sequences of the 18S rDNA deduced from *** were used for designing species-specific primers, and a RFQ-pcr (Real-time fluorescent quantitative Polymerase Chain Reaction) method was developed for quantitative detection of ***. Primer *** and TaqMan probe were designed, and the specificity of primer was checked with pcr. A calibration curve was constructed with cycle threshold value against visual counted cell number. And the value of the curve was tested with other *** samples, which were assayed with both the RFQ-pcr method and visual count under microscope.

关键词： Heterosigma akashiwo fluorescent quantitative pcr molecular probe real-time detection

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Abnormal Expression of Eukaryotic Translation Factors in Malignant Transformed Human Bronchial Epithelial Cells Induced by Crystalline Nickel Sulfide

引用

Biomedical and Environmental Sciences 2006年第1期19卷 53-60页

作者： YI-XIONG LEI XUE-MIN CHEN GEN-RONG WO JIA-KUN CHEN Department of Occupational and Environmental Health Tongji Medical College Huazhong University of Science and Technology Wuhan 430030 Hubei China Department of Preventive Medicine Guangzhou Medical College Guangzhou 510182 Guangdong China

Objective To study the oncogenic potential of mouse translation initiation factor 3 （TIF3） and elongation factor-1δ （TEF-1δ） in malignant transformed human bronchial epithelial cells induced by crystalline nickel sulfide （NiS）. Methods Abnormal expressions of human TIF3 and TEF-1δ genes in two kinds of NiS-transformed cells and NiS-tumorigenic cell lines were investigated and analyzed by the reverse transcript polymerase chain reaction （RT-pcr） and fluorescent quantitative polymerase chain reaction （FQ-pcr）, respectively. Results RT-pcr analysis primarily showed that both human TIF3 and TEF-1δ mRNA expressions in two kinds of NiS-transformed cells and NiS-tumorigenic cell lines were increased as compared with controls. FQ-pcr assay showed that the levels of TIF3 expressions in the transformed cells and tumorigenic cells were 3 and 4 times higher respectively, and the elevated expressions of TEF-16 eDNA copies were 2.7- to 3.5-fold in transformed cells and 4.1- to 5.2-fold in tumorigenic cells when compared with non-transformed cells, indicating that the over-expressions of human TIF3 and TEF-1δ genes were related to malignant degree of the cells induced by nickel. Conclusions These findings demonstrate that there are markedly abnormal expressions of TIF3 and TEF-1δ genes during malignant transformation of human bronchial epithelial cell lines induced by crystalline NiS. They seem to be the molecular mechanisms potentially responsible for human carcinogensis due to nickel.

关键词： Nickel compounds Human cells Transformation Translation factors fluorescent quantitative pcr

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

Nogo-A expression dynamically varies after spinal cord injury

引用

Neural Regeneration Research 2015年第2期10卷 225-229页

作者： Jian-wei Wang Jun-feng Yang Yong Ma Zhen Hua Yang Guo Xiao-lin Gu Ya-feng Zhang Wuxi Hospital of Traditional Chinese Medicine Institute of Orthopedics and Traumatology of Nanjing University of Chinese Medicine Nanjing University of Chinese Medicine

The mechanism involved in neural regeneration after spinal cord injury is unclear. The my-elin-derived protein Nogo-A, which is speciifc to the central nervous system, has been identiifed to negatively affect the cytoskeleton and growth program of axotomized neurons. Studies have shown that Nogo-A exerts immediate and chronic inhibitory effects on neurite *** vivo, inhibitors of Nogo-A have been shown to lead to a marked enhancement of regenerative axon extension. We established a spinal cord injury model in rats using a free-falling weight drop device to subsequently investigate Nogo-A expression. Nogo-A mRNA and protein expression and immunoreactivity were detected in spinal cord tissue using real-time quantitative pcr, immu-nohistochemistry and western blot analysis. At 24 hours after spinal cord injury, Nogo-A protein and mRNA expression was low in the injured group compared with control and sham-operated groups. The levels then continued to drop further and were at their lowest at 3 days, rapidly rose to a peak after 7 days, and then gradually declined again after 14 days. These changes were observed at both the mRNA and protein level. The transient decrease observed early after injury followed by high levels for a few days indicates Nogo-A expression is time dependent. This may contribute to the lack of regeneration in the central nervous system after spinal cord injury. The dynamic varia-tion of Nogo-A should be taken into account in the treatment of spinal cord injury.

关键词： nerve regeneration spinal cord injury contusion Nogo-A axon growth immunohisto-chemistry fluorescent quantitative pcr neural regeneration

来源：

维普期刊数据库

同方期刊数据库评论

在线全文

学校读者我要写书评

暂无评论

欢迎您,

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

请选择保存的检索档案：

请选择收藏分类：

通借通还

欢迎您,

建议与咨询 留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

时间限定

文献类型

馆藏选择

核心期刊

语言

文献类型

帮助

文字说明：

检索规则说明：

检索范例：

分类表

所选分类

限定检索结果

文献类型

馆藏范围

日期分布

学科分类号

主题

机构

作者

语言

在线全文

在线全文

在线全文

在线全文

在线全文

在线全文

请选择保存的检索档案： 新增检索档案 确定 取消

请选择收藏分类： 新增自定义分类 确定 取消

通借通还

建议与咨询留下您的常用邮箱和电话号码，以便我们向您反馈解决方案和替代方法

请选择保存的检索档案：

请选择收藏分类：