许文涛
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- 姓名:许文涛
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学科领域:
食品科学技术
- 研究兴趣:
许文涛,男,1979年出生于安徽省阜阳市,博士,一级讲师。
主要研究领域:转基因产品食用安全评价技术与检测技术、食品安全风险评估技术与检测技术。
主要学术和行政兼职:农业部农业转基因生物食用安全监督检验测试中心(北京)主任助理兼分子检测室主任;农业部农产品质量监督检验测试中心(北京)重金属和微生物检测室主任。
主要研究经历:2001——2006年,中国农业大学,硕博连读研究生,导师为罗云波教授,主要研究转基因产品食用安全和转基因成分检测技术;期间赴加拿大学习和交流。主要参与和主持了农业部“948”项目(两项)、科技部“863”项目(两项)以及其他10余项国家级以及省部级课题的研究。目前已经获得教育部成果鉴定一项,已申请国家专利10项,其中授权国家专利3项,参与制定转基因领域国家标准一项,主编北京市精品教材《转基因食品安全检测与检测技术》。并且在食品安全领域已经发表40余篇学术论文,其中十余篇文章被SCI以及EI收录。发表杂志涉及:《Journal of Agriculture and Food Chemistry》、《Journal of the Science of Food and Agriculture》、《Food control》、《Food Science》、《Plant Cell Repots》、《Biotechnol Lett》、《Meat Science》、《Journal of Food Biochemistry》、《Food Chemistry》、《Protein Expression and Purification》、《Chinese Journal of Agricultural biotechnology》、《Meat Science》、《Postharvest Biology and Technology》《农业生物技术学报》、《食品科学》、《农业科技导报》等。
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11
许文涛, Wen-Tao Xu, Kun-Lun Huang, Ying Wang, Hong-Xing Zhang and Yun-Bo Luo
Journal of the Science of Food and Agriculture J Sci Food Agric 86:1103–1109 (2006),-0001,():
-1年11月30日
Biotechnology has permitted the modification of agricultural materials in a very precise way to improve productivity and yields. Polymerase chain reaction (PCR)-based methods have been the first choice of most analytical laboratories for routine use in the detection of genetically modified organisms (GMO) and their derived products. These methods rely on the amplification of transgenic sequences and quantification of the transgenic DNA by comparison with an amplified reference gene. This paper describes the specific primers and probe for the cotton stearoyl-ACP desaturase (sad1) gene, and PCR cycling conditions suitable for the use of this sequence, which acts as an endogenous reference gene in both qualitative and quantitative PCR assays. The two methods were tested with 18 cotton varieties and identical amplification products were obtained with all of them. No amplification products were detected when DNA samples from other species, including soybean, rapeseed, tobacco, maize, tomato, potato, cucumber, pea, red pepper, sunflower, sesame, rice, peach, banana, apple, pumpkin, barley and carrot, were used as templates, which demonstrates that this system is specific for cotton. In real-time quantitative PCR analysis, the detection limit was as low as 6 pg of DNA, which indicates that this method is suitable for application to processed food samples that contain very low copies of target DNA. Southern blot analysis confirmed that the sad1 gene was a single copy in the tested cotton varieties. 2006 Society of Chemical Industry
cotton, stearoyl-ACP desaturase, sad1 gene, polymerase chain reaction, genetically modified organisms
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许文涛, Wentao Xu, Wei Qu, Kunlun Huang, Feng Guo, Jiajia Yang, Heng Zhao, YunBo Luo
Postharvest Biology and Technology 45 (2007) 126–133,-0001,():
-1年11月30日
The application of Grapefruit Seed Extract (GSE) as a sanitizer for reducing the populations of human bacterial pathogens on whole and fresh-cut green vegetables was investigated. Cucumber and lettuce were selected as model green vegetables, and six bacteria strains, including three strains of Salmonella spp. and three strains of Listeria monocytogenes, were selected in our study in order to determine the antibacterial effects of sanitizers. The survival and growth of total aerobes, Salmonella spp., and L. monocytogenes on whole and fresh-cut cucumber and lettuce during storage (10 and 4℃) were analyzed by using a classical microbiological enumeration. The antibacterial effects of GSE alone and GSE in combination with nisin and citric acid (Mixture agent II) were significant (P < 0.05). Treatments with sodium lactate and potassium sorbate (Mixture agent I) were also tested as chemically synthesized agents. Sensory quality was evaluated, and there was no significant difference between GSE and Mixture agent II treatments during storage in terms of organoleptic and visual properties. Our results suggest that GSE could inhibit bacteria significantly (P < 0.05) and prolong the preservation time; GSE might be applied as an effective and safe preservative for ready-to-eat cucumber and lettuce.
Grapefruit Seed Extract, Salmonella, Listeria, Sensory quality, Cucumber, Lettuce
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许文涛, WENTAO XU, KUNLUN HUANG, HENG ZHAO, , AND YUNBO LUO
J. Agric. Food Chem. 2005, 53, 4315-4321,-0001,():
-1年11月30日
We have developed a new immunoassay method to detect genetically modified (GM) maize and rape containing phosphinothricin-N-acetyltransferase (PAT). PAT encoded by Bialaphos resistance gene (bar) was highly expressed in soluble form in Escherichia coli BL21(DE3) and purified to homogeneity by Ni2+ affinity chromatography. A simple and efficient extraction and purification procedure of PAT from GM maize and rape was developed by means of the immunoaffinity column (IAC) as a cleanup tool. Purified polyclonal antibodies against PAT was produced and coupled covalently to CNBr-activated Sepharose 4B. Both the binding conditions and elution protocols were optimized. The IAC was successfully employed to isolate and purify the PAT from the various tissues of GM maize (Bt11 and Bt176) and rapes (MS1/RF1 and MS8/RF3). Enzyme linked immunosorbent assay (ELISA) procedures were established further on to measure the PAT protein. GM maize cannot be differentiated from non-GM maize by ELISA. But IAC-ELISA allowed 0.5% GMOs to be detected in MS1/RF1 and MS8/RF3 and 10% GMOs to be detected in Bt11 and Bt176, which makes this method an acceptable method to access PAT protein in GM rapes and maize.
Purification, PAT, GMO, detection, immunoaffinity column, enzyme linked immunosorbent assay
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许文涛, Wen-Tao Xu, , Kun-Lun Huang, Ai-Ke Deng, Zhi-hong Liang, Yun-Bo Luo
Food Control 18 (2007) 1300–1306,-0001,():
-1年11月30日
The present study investigated the DNA density of the embryo, cotyledon, and seed coat of each soybean from 15 soybean cultivars, and evaluated the impacts of variations of tissue DNA density and nuclear DNA content in soybean lines on GMO quantification. The results have shown that DNA densities and DNA quantity ratios among the various tissues of soybean are significantly different from each other and have insignificant influence on the transgenic copy number and therefore on GMO quantification. The nature of the CRM and the transgenic material present in the analyzed sample has prominent impact on transgenic genome copies, thus on GMO quantification. The DNA densities of the various soybean-derived products are also distinctly different from that of the whole soybean, but which does not result in the similar differences on the GMO ratio among them. Nuclear DNA content of soybean is different from cultivar to cultivar. Results shows that variation of nuclear DNA content in soybean lines has a great impact on the accurate determination of GMO. In some extreme situations, the deviation amplitude can reach 26%, which is intolerable for accurate determination. _ 2006 Published by Elsevier Ltd.
DNA density, Nuclear DNA content, GMO, Soybean, Impact, Quantification
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许文涛, Rong Yang, Wentao Xu, Yunbo Luo, Feng Guo, Yun Lu, Kunlun Huang,
Plant Cell Rep (2007) 26:1821–1831,-0001,():
-1年11月30日
With the development of genetically modified organisms, labeling regulations have been introduced, which require appropriate detection methods. Event-specific qualitative and quantitative polymerase chain reaction (PCR) detection methods have become the internationally agreed state-of-art. This paper describes an event-specific PCR method for qualitative and quantitative of Roundup Ready canola event GT73. The 3¢-integration junction was characterized by two methods: inverse-PCR and thermal asymmetric interlaced-PCR. In the conventional qualitative PCR assay, the event-specific primers designed were confirmed to be specific and the limit of detection (LOD) was 0.05% (approximates to ten haploid genome copies). In the quantitative TaqMan real-time PCR assay, the LOD and the limit of quantification were five and ten haploid genome copies, respectively. In addition, for further quantitative detection, a reference molecule which contained the canola endogenous gene and event-specific sequence was constructed and standard curves were set up. The goodness of the linearity and high efficiency of the PCR reaction indicated the usability of the plasmid and the established PCR system. Moreover, mixed samples with different GT73 content (6, 3, 1 and 0.5%) were quantified using the established real-time PCR system to evaluate the trueness and precision of the system. The trueness expressed as bias varied from 2.00 to 18.00%. The precision expressed as variation coefficient were different from 6.40 to 32.95%. From above results, we believed that the established eventspecific qualitative and quantitative PCR systems for GT73 in this study were acceptable and suitable for genetic modified canola detection.
Event-specific, Roundup ready canola event GT73 (, Brassica napus), , Integration junction, Inverse PCR, Thermal asymmetric interlaced, TaqMan real-time PCR
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许文涛, Y. LU, W. XU, , A. KANG, Y. LUO, F. GUO, R. YANG, J. ZHANG, AND K. HUANG
JFS M: Food Microbiology and Safety Vol. 72, Nr. 7, 2007—JOURNAL OF FOOD SCIENCE,-0001,():
-1年11月30日
The hygromycin B phosphotransferase gene (hpt) has been widely used in the process of plant genetic engineering to produce plants that can secrete the HPT protein. As part of a safety assessment, sufficient quantities of the proteinwere produced in Escherichia coli to conduct in vitro digestibility and animal studies.Western blotting analysis showed that the HPT protein was digested by simulated gastric fluid within 40 s. ELISA demonstrated that the protein did not induce detectable levels of specific IgE antibodies or histamine in test animals. Alignment of the amino acid sequence of HPT with those of known allergens did not produce evidence of sequence similarities between these allergens and the HPT protein.We conclude that HPT has a low probability to induce allergenicity.
allergenicity,, animalmodel,, HPT protein,, in vitro digestibility,, safety assessment
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许文涛, Wen-Tao Xu, , Kun-Lun Huang, Feng Guo, Wei Qu, Jia-Jia Yang, Zhi-Hong Liang, Yun-Bo Luo
Postharvest Biology and Technology 46 (2007) 86–94,-0001,():
-1年11月30日
‘Redglobe’ table grapes (Vitis vinifera cv. Redglobe), undergoing deterioration were selected as model fruit with, Botrytis cinerea, to test the antifungal activity of grapefruit seed extract (GSE) in vitro and in vivo. The results of inhibition of spore germination and radial growth of B. cinerea in vitro indicated that GSE could efficiently inhibit the growth of the tested fungi. The effectiveness of GSE and chitosan to control postharvest decay and quality of ‘Redglobe’ grape berries stored at 0–1 ◦Cwas also investigated. Chitosan and GSE treatments, alone or combined, significantly reduced postharvest fungal rot of the fruit compared with controls challenged with B. cinerea. Differences in weight loss, color change, ripening, sensory quality and microorganism index between grapes treated with GSE and control fruit suggested that GSE had both antifungal and antioxidative activity. Moreover, the sensory analyses revealed beneficial effects in terms of delaying rachis browning and dehydration and maintenance of the visual aspect of the berry without detrimental effects on taste, or flavour. GSE and chitosan might have a synergistic effect in reducing postharvest fungal rot and maintaining the keeping quality of ‘Redglobe’ grapes.
Grapefruit seed extract, Chitosan, Botrytis cinerea, Table grape, Quality attributes
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许文涛, Wentao Xu, Kunlun Huang, Aike Deng, Baiqiang Zhai, Heng Zhao, , Yingcong Li, Zhihong Liang, Yunbo Luo
African Journal of Food Science. pp. xxx-xxx,October, 2007,-0001,():
-1年11月30日
Ractopamine has been developed to be the main β-agonist substance used illegally in meat producing animals. A simple and efficient extraction and purification procedure for ractopamine was developed by means of the immunoaffinity column (IAC) as a cleanup tool. Purified polyclonal antibodies against RCT were produced and coupled covalently to CNBr-activated Sepharose 4B. Both the binding conditions and elution protocols were optimized and the capacity, reusability, precision and accuracy of IAC were determined. The IAC was successfully employed to isolate and purify the RCT from the various tissues of swine. Subsequently, enzyme linked immunosorbent assay (ELISA) procedures were established further on to measure RCT. The antibodies showed negligible cross-reactivity with other β-agonists. IAC-ELISA allowed 0.2 ng/mL of RCT to be detected in urine and 0.5 ng/mL to be detected in other various tissues of swine, which makes this method an acceptable screening tool to access RCT. IACELISA for the detection of RCT was validated by LC-MS and the correlations between the results from LC-MS and those from IAC-ELISA were all high (above 0.89).
Purification,, ractopamine,, swine,, immunoaffinity column,, enzyme linked immunosorbent assay
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许文涛, Xijin Lin, Wentao Xu, , Kunlun Huang, Xiaohong Mei, Zhihong Liang, Zhemin Li, Jingxin Guo, YunBo Luo
,-0001,():
-1年11月30日
The gene lasB from Pseudomonas aeruginosa, which encoded elastase, was cloned and firstly successfully expressed in Pichia pastoris stain KM71 under the control of AOX promoter. The effects on the recombinant elastase activities of different pH, different temperatures and different metal ions were assayed. The full-length gene (1497 bp) encodes a preproenzyme including a N-terminal signal peptide (23 aa), a propeptide (197 aa) and mature elastase (301 aa). The recombinant elastase was secreted into culture supernatants using signal sequence from lasB and showed a single band at about 34 kDa by SDS-PAGE. The recombinant elastase expression hit the highest level of approximately 450 mg/L and the specific elastolytic activity of the recombinant elastase was 130 U/ml, which was approximately 26-fold higher than that of elastase obtained from P. aeruginosa. The optimal temperature and pH of the recombinant elastase was 28℃ and 7.4, respectively. The enzyme possessed high resistance to heat, and can be activated by Ca2+. These enzyme properties suggested that it could be produced in an industrial scale and has the potential to be a commercial enzyme.
recombinant Elastase, Picha pastoris, Pseudomonas aeruginosa
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【期刊论文】稻米深加工产品基因组提取方法及其对PCR 的影响*
许文涛, , 黄昆仑, 芦云, 郭峰, 杨蓉, 秦伟, 罗云波
农业生物技术学报/Journal of Agricultural Biotechnology 2007, 15 (1): 97~101,-0001,():
-1年11月30日
以4 种米粉为原料,每个样品做3个梯度(120、800 和2000mg),采用改进的经典酚/仿法、CTAB(十六烷基三乙基溴化铵)沉淀法以及盐酸胍/氯仿法提取基因组,经PCR 扩增内标基因(SPS)检测方法的优劣。结果显示,120 mg 的样品经3 种方法提取的基因组均不能扩增出内标基因;800和2000mg 的样品只有用CTAB沉淀法提取的基因组(采用相同的模板量)能全部扩增出内标基因。结果表明,CTAB 沉淀法提取基因组的效果最好,对PCR的抑制现象最少。
稻米深加工产品, 基因组, PCR, 十六烷基三乙基溴化铵沉淀法
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