Chinese Journal of Agricultural Biotechnology最新文献_第4页

Establishment and evaluation of real-time PCR for West Nile virus detection. 西尼罗病毒实时PCR检测方法的建立及评价。

Chinese Journal of Agricultural Biotechnology Pub Date : 2009-04-01 DOI: 10.1017/S1479236209002599

Shi Lijun, Lü MaoMin, L. Gang, Li Cheng-yao, Zhang Jingang

引用次数: 4

Real-time PCR assay for sensitive organ detection and epidemic investigation of Turbot reddish body iridovirus 大菱鲆红体虹膜病毒敏感器官的实时荧光定量PCR检测及流行病学调查

Chinese Journal of Agricultural Biotechnology Pub Date : 2009-04-01 DOI: 10.1017/S1479236209002605

Wu Cheng-long, Shi Cheng-yin, H. Jie, Kong Xiaoyu

{"title":"Real-time PCR assay for sensitive organ detection and epidemic investigation of Turbot reddish body iridovirus","authors":"Wu Cheng-long, Shi Cheng-yin, H. Jie, Kong Xiaoyu","doi":"10.1017/S1479236209002605","DOIUrl":"https://doi.org/10.1017/S1479236209002605","url":null,"abstract":"A rapid and sensitive Real-time PCR assay coupled with SYBR Green Ⅰchemistry was developed for the quantitative detection of Turbot reddish body iridovirus (TRBIV) isolated from farmed turbot (Scophthalmus maximus). A 152 bp DNA fragment from TRBIV major capsid protein (MCP) gene was involved in the Real-time PCR assay using the Roter Gene 3000 sequence detection system. The PCR mixture contained a fluorescence dye, SYBR Green Ⅰ, which binding to dsDNA exhibited fluorescence enhancement. The enhancement of fluorescence was proportional to the initial concentration of the template DNA. The positive control plasmid pUCm-T/TRBIV MCP containing the target sequence was quantified to make the standard curve for sample detection after serial 10-fold dilution. Linear coefficient correlations between cycle threshold (CT) value and logarithmic positive plasmid concentration were close to one (r2=0.9952) and the detection limit of the assay was 102 copies of positive plasmids. The quantitative detection of different tissues from TRBIV-infected fish showed that the spleen and kidney contained the largest number of viral particles (5.23×106 and 2.18×106 viral genome copies/mg tissue, respectively) while no viral DNA was detected in the muscular tissue. The molecular epidemic investigation of TRBIV showed that many cultured turbots were infected and TRBIV has been spread widely in turbot farms locating at Shandong peninsula. The virus number was varied from 1.27×102 to 2.33×106 of viral genome copies/mg tissue in spleen of infected turbot. These results suggest that the Real-time PCR assay reported here can be used as a rapid, sensitive and quantitative method for TRBIV.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"21 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129382699","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

Expression optimization of subunit fasG of 987P fimbriae in enterotoxigenic Escherichia coli (ETEC) 987P菌膜fasG亚基在产肠毒素大肠杆菌(ETEC)中的表达优化

Chinese Journal of Agricultural Biotechnology Pub Date : 2009-04-01 DOI: 10.1017/S1479236209002551

Lu He-feng, Wang Jie, Gou Zhong-Yong, Peng Jian

引用次数: 1

Construction and utilization of plant expression vectors with PMI gene as selection marker to recover transgenic plants of Citrus sinensis L. Osbeck 以PMI基因为选择标记的柑橘植物表达载体的构建与利用

Chinese Journal of Agricultural Biotechnology Pub Date : 2009-04-01 DOI: 10.1017/S1479236209002678

Z. Li-hui, Xuan Hai-feng, Wang Hui-Quang, W. Shaohua, Zhuang Yi-xuan

引用次数: 1

Growth and identification of human amniotic fluid stem cells and analysis of their influencing factors 人羊水干细胞的生长鉴定及其影响因素分析

Chinese Journal of Agricultural Biotechnology Pub Date : 2009-04-01 DOI: 10.1017/S1479236209002630

W. Han, Dou Zhong-ying, Wang Hua-yan

{"title":"Growth and identification of human amniotic fluid stem cells and analysis of their influencing factors","authors":"W. Han, Dou Zhong-ying, Wang Hua-yan","doi":"10.1017/S1479236209002630","DOIUrl":"https://doi.org/10.1017/S1479236209002630","url":null,"abstract":"Amniotic fluid stem (AFS) cells derived from human amniotic fluid were cultured in vitro and the factors of affecting the primary culture of human AFS cells were investigated. The isolated AFS cells, which express embryonic stem cell markers, such as Oct-4, and adult stem cell markers, such as CD29, grew faster and had high proliferation ability. Moreover, AFS cells could differentiate into neurons and cardiomyocytes which expressed Nestin and α-actin, respectively. There were several factors, such as the date of gestation, the level of blood contamination and the volume of amniotic fluid, could significantly affect the attachment time and numbers of AFS cells in the primary culture. The results showed that cell attachment time of the 2nd trimester of gestation (4.7±0.6) d was significantly different from that of the 3rd trimester of gestation (6±0.5) d (P 0.05), suggesting that cells collected from the fluid of 3rd trimester of gestation needed longer attachment time. Blood contamination could significantly affect the cell attachment time. The attachment time of brown-color fluid group (10.8±0.3) d was significantly different with the blood-cell-free fluid group (6±0.5) d and blood-cell group (6.3±0.6) d (P 0.05). The volume of amniotic fluid influenced on cell attachment numbers and time to some extent. With the increase of amniotic fluid volume, cell attachment numbers significantly increased (P 0.05), and cell attachment time extended but no significant difference(P 0.05). The present studies systematically examined the factors, effection on primary culture of human AFS cells and provided some useful data for AFS cell research. Additionally, the isolated AFS cells maintain the capabilities of differentiation into other cell types and are able to become seed cells for the clinical application.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"1 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"130796325","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1

In vitro effect of lentinan on the activation of immunological cells in Cyprinus carpio 香菇多糖对鲤免疫细胞活化的体外影响

Chinese Journal of Agricultural Biotechnology Pub Date : 2009-04-01 DOI: 10.1017/S1479236209002496

Zhang Liu, Cao Liping, Ding Weidong, Galina Jeney, X. Pao, Xiong Chuan-xi, Yin Guojun

引用次数: 4

Expressed sequence tag analysis and cDNA array establishment of Nicotiana tabacum : application to salinity stress 烟草表达序列标记分析及cDNA阵列的建立及其在盐胁迫中的应用

Chinese Journal of Agricultural Biotechnology Pub Date : 2009-04-01 DOI: 10.1017/S1479236209002526

Li Wenzheng, Song Li-min, Li Yongping, L. Xiuping, Lu Hongmei, Dai Chengen, Fang Yongqi, Dong Hai-tao, L. Debao

引用次数: 3

Construction of a membrane-lytic immunotoxin using melittin and epidermal growth factor 利用蜂毒素和表皮生长因子构建溶膜免疫毒素

Chinese Journal of Agricultural Biotechnology Pub Date : 2009-04-01 DOI: 10.1017/S1479236208002362

Chen Mei-ling, N. Dong, Ruan Hui, Pan Bing-qing, Wan Jin-ling, Wu De, Zhang Jia-jia, Chen Qihe, He Guoqing

{"title":"Construction of a membrane-lytic immunotoxin using melittin and epidermal growth factor","authors":"Chen Mei-ling, N. Dong, Ruan Hui, Pan Bing-qing, Wan Jin-ling, Wu De, Zhang Jia-jia, Chen Qihe, He Guoqing","doi":"10.1017/S1479236208002362","DOIUrl":"https://doi.org/10.1017/S1479236208002362","url":null,"abstract":"Epidermal growth factor receptor (EGFR) is becoming a perfect target for killing carcinoma cells, especially because of its overexpression on the surface of these cells. Cationic antimicrobial peptides (CAP) have their own special mechanism of membrane-lytic cytotoxicity. In this study, a membrane-lytic immunotoxin (IT), chimeric protein MEGFMEL, was constructed to kill carcinoma cells with EGFR overexpression. This protein is composed of mouse ( Mus musculus ) epidermal growth factor (MEGF), as the target part, and melittin (MEL), as the cytotoxic part. Using Escherichia coli BL21 and pET30a as expression strain and vector, respectively, 63.45 μg/ml of MEGFMEL (68% purity) was obtained through low-temperature induction of expression and a thawing-freezing purification procedure (without cytolysis). In vitro activity measurement showed that this MEGFMEL significantly induced a lethal effect on A 431 carcinoma cells overexpressing EGFR on the surface, with an LD 50 value 52.6 μg/ml. The results suggest that the use of CAP as the toxin in the construction of unique membrane-lytic ITs aimed at EGFR is feasible.","PeriodicalId":236932,"journal":{"name":"Chinese Journal of Agricultural Biotechnology","volume":"25 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2009-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"133649584","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Research progress in techniques for detecting genetically modified organisms 转基因生物检测技术的研究进展

Chinese Journal of Agricultural Biotechnology Pub Date : 2009-04-01 DOI: 10.1017/S1479236209002575

Xu Wentao, Bai Wei-bin, Luo Yunbo, Yuan Yanfang, Huang Kunlun

引用次数: 9

Auxin regulation in the interaction between Rice stripe virus and rice. 水稻条纹病毒与水稻互作中生长素的调控。

Chinese Journal of Agricultural Biotechnology Pub Date : 2009-04-01 DOI: 10.1017/S1479236209002502

Yang Jin-guang, Wang Wen-ting, Ding Xin-Lun, Guo Li-juan, Fang Zhen-Xing, Xie Liyan, Lin Qi-ying, Wu Zujian, Xie Lian-hui

引用次数: 2