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1.
人乳铁蛋白腺病毒载体的构建及细胞表达 总被引:1,自引:0,他引:1
将人乳铁蛋白基因与腺病毒基因组骨架质粒重组,在293细胞中包装重组腺病毒颗粒,获得真核细胞表达的栽体pAd-hLTF.以pcDNA3X为模板,PCR扩增hLTF基因,腺病毒穿梭栽体pshuttle-cmv与hLTF重组为pshuttle-cmv-hLTF:再与腺病毒骨架质粒pAdEasy-1同源重组为pAd-hLTF质粒:脂质体法将重组pAd-hLTF质粒转染HEK 293细胞.包装成重组腺病毒颗粒.Western blot和ELISA法测定细胞中hLTF基因的表达.结果表明,pAd-hLTT质粒转染293细胞后,7~10 d后,获得细胞裂解液,将细胞裂解产物再次接种新鲜的293细胞,3~5 d后,80%以上的细胞变圆、膨胀、漂浮.Westem blot和ELISA结果显示,上清液中有hLTF基因的表达,为重组人乳铁蛋白的体外表达及其功能分析奠定了基础. 相似文献
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由于DNA重组技术和蛋白组技术的发展与成熟,外源基因表达技术备受关注。其在生物、食品、工业以及医学领域发挥着举足轻重的作用。以原核生物细胞及真核生物细胞作为表达系统,进行目的基因序列的表达以生产蛋白疫苗、核酸疫苗和生物酶制剂等是近年来发酵工业的新型领域。原核和真核表达系统是近年来研究和应用最广泛和最成熟的外源基因表达系统。对近年来关于大肠杆菌表达系统和酵母表达系统的研究进展进行了综述。 相似文献
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S.H.P.P. Karunaratne N.J. Hawkes H. Ranson 《Pesticide biochemistry and physiology》2007,88(1):108-113
The present status of pyrethroid resistance in vectors of malaria; Anopheles culicifacies and Anopheles subpictus, was tested in two malarious Districts, Anuradhapura and Trincomalee, of Sri Lanka. Both species were resistant to permethrin and susceptible to cypermethrin and cyfluthrin. An. subpictus were resistant to deltamethrin. λ-Cyhalothrin and etofenprox resistance was shown only by Anuradhapura An. subpictus. Although there were no differences among the populations for esterase and glutathione S-transferase activities, increased monooxygenase levels were found among Trincomalee populations. The voltage-gated sodium channel gene, the target site gene of pyrethroids, was partially sequenced to screen for mutations previously associated with insecticide resistance. The classic leucine to phenylalanine substitution, TTA to TTT, was detected in An. subpictus. It appears that both kdr type and monooxygenase resistance underlie pyrethroid resistance in these two malaria vectors of Sri Lanka. 相似文献
6.
Hsiu-Lin Liu Ching-Chung Chen Chan-Pin Lin 《European journal of plant pathology / European Foundation for Plant Pathology》2007,117(3):281-291
Pear decline (PD) is an important phytoplasmal disease that occurs mainly in Europe and North America. In 1994, pear trees
exhibiting symptoms typical of PD disease were observed in orchards of central Taiwan. The sequence of 16S rDNA and 16S–23S
rDNA intergenic spacer region (ISR) of the causative agent of pear decline in Taiwan (PDTW) were amplified with polymerase
chain reaction (PCR) using a DNA template prepared from the diseased leaves. Sequence analysis of 16S rDNA revealed that the
PDTW agent was closely related to the phytoplasmas of the apple proliferation group that cause diseases in stone fruits, pear
and apple. Consistent with the result of 16S rDNA sequence analysis, sequence analysis of the 16S–23S rDNA ISR and putative
restriction site analyses of 16S rDNA and 16S–23S rDNA ISR sequences provided further support for the view that the PDTW phytoplasma
causing pear decline in Taiwan may represent a new subgroup of the apple proliferation group. According to the rDNA sequence
of PDTW phytoplasma, two specific PCR primer pairs, APf2/L1n and fPD1/rPDS1, were designed in this study for the detection
of the etiological agent in pear trees and insect vectors. Based on the sequence analyses of the PCR-amplified fragments,
two species of pear psyllas, Cacopsylla qianli and Cacopsylla chinensis, were found to carry PDTW phytoplasma. 相似文献
7.
麦二叉蚜传播玉米矮花叶病毒的机制 总被引:2,自引:0,他引:2
用胶体金标记法和荧光抗体标记法研究了麦二叉蚜(Schizaphis graminum)传播玉米矮花叶病毒(Maize dwaft mosaic virus,MDMV)的机制。麦二叉蚜传播玉米矮花叶病毒需要辅助成份-蛋白酶(Helper component-proteinase,HC-Pro)的参与。在电镜下观察到HC-Pro可以与MDMV粒子结合。用FITC标记的HC-Pro抗体和MDMV抗体证明,HC-Pro可以直接结合到蚜虫口针上;而MDMV粒子不能直接结合到蚜虫口针,必须在HC-Pro的辅助下才能结合到蚜虫口针上。这为HC-Pro在蚜虫传毒过程中起桥梁作用提供了新的证据。MDMV粒子主要吸附在蚜虫口针的尖端和中间部分。 相似文献
8.
Humberto J.O. Ramos M. Geoffrey Yates Emanuel M. Souza 《Soil biology & biochemistry》2011,43(8):1626-1638
Bacteria are extraordinarily diverse microorganisms with a huge potential to benefit environmental and agricultural systems. Comprehensive studies in complex habitats such as soils and plants have led to the development of genetic tools to evaluate gene expression and bacterial colonization under controlled or environmental conditions and to obtain genetically engineered organisms for environmental release. In addition, current advances in genomic and metagenomic research will add to the number of genes with potential for biotechnological applications, which will require the development of appropriate genetic systems to fulfill their potential for both industrial and agricultural applications. The aim of the present review is to assess the approaches and recent progress in vector design and genetic tools to study and manipulate plant-bacterial interactions, as well as strategies to construct genetically modified strains for environmental release. 相似文献
9.
Hyeshin Hwang Younghye Ro Hyunkyoung Lee Jihyeon Kim Kyunghyun Lee Eun-Jin Choi You-Chan Bae ByungJae So Dohoon Kwon Ho Kim Inhyung Lee 《Journal of veterinary science (Suw?n-si, Korea)》2022,23(5)
BackgroundSince 2013, the number of requests for diagnosis for horses based on neurological symptoms has increased rapidly in South Korea. The affected horses have commonly exhibited symptoms of acute seasonal hindlimb ataxia. A previous study from 2015–2016 identified Setaria digitata as the causative agent.ObjectivesThis study is an epidemiological investigation to find out risk factors related to the rapid increase in hindlimb ataxia of horses due to aberrant parasites in South Korea.MethodsAn epidemiological investigation was conducted on 155 cohabiting horses in 41 horse ranches where the disease occurred. The surrounding environment was investigated at the disease-causing horse ranches (n = 41) and 20, randomly selected, non-infected ranches.ResultsHindlimb ataxia was confirmed in nine cohabiting horses; this was presumed to be caused by ectopic parasitism. Environments that mosquitoes inhabit, such as paddy fields within 2 km and less than 0.5 km from a river, had the greatest association with disease occurrence.ConclusionsMost horse ranches in South Korea are situated in favorable environments for mosquitoes. Moreover, the number of mosquitoes in the country has increased since 2013 due to climate change. Additional research is required; however, these data show that it is necessary to establish guidelines for the use of anthelmintic agents based on local factors in South Korea and disinfection of the environment to prevent disease outbreaks. 相似文献
10.
用PCR的方法,从产甘油假丝酵母WL2002—5中扩增出了产甘油关键酶3-磷酸甘油脱氢酶(glycerol 3-phosphate dehydrogenase)和3-磷酸甘油磷酸酶(glycerol 3-phosphate phosphatase)的基因GPD和GPP。对这两个基因测序,发现基与酿酒酵母(5accharomyces.cereuisiae)的GPD、GPP基因的相似性达99%。并构建了其在植物中的表达载体。 相似文献