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抗Spodoptera f.病毒单克隆抗体的初步研制
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作者 陆苹 Michael Ma 《上海交通大学学报(农业科学版)》 1987年第1期1-5,共5页
本文介绍了 Spodoptera frugiperda 病毒多角体及病毒粒子的提纯方法。粗提材料先通过40%蔗糖,在10000r/min 下离心15分钟,随后悬浮于0.01M Tris 缓冲液,25~65%蔗糖的连续梯度中,于40000 r/min 下离心30分钟。取出多角体的条带,洗除游... 本文介绍了 Spodoptera frugiperda 病毒多角体及病毒粒子的提纯方法。粗提材料先通过40%蔗糖,在10000r/min 下离心15分钟,随后悬浮于0.01M Tris 缓冲液,25~65%蔗糖的连续梯度中,于40000 r/min 下离心30分钟。取出多角体的条带,洗除游离残存的蔗糖,透析过夜。将多角体在4℃,0.01MNa_2CO_3,0.01MEDTA,0.17M NaCl,pH=1下,作用1小时,随后调节 pH 到8.5~8.9,2000g,离心10分钟。该制剂在4℃下,25~60%蔗糖梯度中,24000r/min,离心2小时。随后取出病毒粒子,重新悬浮于蒸馏水中,于—70℃下贮藏备用。每个 BALB/C 小白鼠免疫4~5次,最后一次注射是在融合前4~7天。将2×10~8脾细胞与2×10~7P_3×63或 SP-2骨髓瘤细胞混合,并用50%PEG(Sigma)融合,用间接酶联免疫法进行筛选阳性杂交瘤细胞,强阳性者进行克隆化。以多角体及病毒粒子为抗原者,均建立了10株能产生特异性单克隆抗体的杂交瘤细胞株,该抗体的效价较高,并进行了血清学的分类。 展开更多
关键词 SPODOPTERA f.病毒 多角体 单克隆抗体
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Cloning and Sequence Analysis of HN and F Protein Genes from a Strain of Goose Paramyxovirus 被引量:2
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作者 易春华 潘杰 +3 位作者 付薇 颜健华 徐贤坤 熊毅 《Agricultural Science & Technology》 CAS 2009年第4期75-78,共4页
[ Objective] The study was to clone HN and F genes from GX1 strain of goose paramyxovirus and analyze their sequences. [ Method] According to the full nucleotide sequence of GPV-SF02 strain of goose paramyxovirus, two... [ Objective] The study was to clone HN and F genes from GX1 strain of goose paramyxovirus and analyze their sequences. [ Method] According to the full nucleotide sequence of GPV-SF02 strain of goose paramyxovirus, two pairs of pdmers were designed to amplify the HN and F genes from GX1 strain of goose paramyxovirus isolated from diseased goose in Guangxi Zhuang Autonomous Region; the amplified products were ligated into pMD18-T vector and sequenced. [ Result ] HN and F genes of this strain tested were 1 716 and 1 662 bp in full nucleotide length, respectively; both showed the homologues of about 97.3% with GPV- SF02 strain, of 80.3% -97.5% with strains LaSota, F48E9 and JS, of just 84.8% with Miyadera strain. [ Conclusion] The results show that isolated strain BX1 matches to virulent APMV-1 strain, belonging to genotype Ⅶ of APMV-1 strain. 展开更多
关键词 Goose paramyxovirus HN protein gene f protein gene CLONING Sequence analysis
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Classifying genotype F of hepatitis B virus into F1 and F2 subtypes 被引量:6
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作者 Hideaki Kato Kei Fujiwara +9 位作者 Robert G. Gish Hiroshi Sakugawa Hiroshi Yoshizawa Fuminaka Sugauchi Etsuro Orito Ryuzo Ueda Yasuhito Tanaka Takanobu Kato Yuzo Miyakawa Masashi Mizokami 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第40期6295-6304,共10页
AIM: To explore the propriety of providing hepatitis B virus (HBV) genotypes F and H with two distinct genotypes. METHODS: Eleven HBV isolates of genotype F (HBV/F) were recovered from patients living in San Fra... AIM: To explore the propriety of providing hepatitis B virus (HBV) genotypes F and H with two distinct genotypes. METHODS: Eleven HBV isolates of genotype F (HBV/F) were recovered from patients living in San Francisco, Japan, Panama, and Venezuela, and their full-length sequences were determined. Phylogenetic analysis was carded out among them along with HBV isolates previously reported. RESULTS: Seven of them clustered with reported HBV/F Isolates in the phylogenetic tree constructed on the entire genomic sequence. The remaining four flocked on another branch along with three HBV isolates formerly reported as genotype H. These seven HBV isolates, including the four in this study and the three reported, had a sequence divergence of 7.3-9.5% from the other HBV/F isolates, and differed by 〉13.7% from HBV isolates of the other six genotypes (A-E and G). Based on a marked genomic divergence, falling just short of 〉8% separating the seven genotypes, these seven HBV/F isolates were classified into F2 subtype and the former seven into F1 subtype provisionally. In a pairwise comparison of the S-gene sequences among the 7 HBV/F2 isolates and against 47 HBV/F1 isolates as well as 136 representing the other six genotypes (A-E and G), two clusters separated by distinct genetic distances emerged.CONCLUSION: Based on these analyses, dassifying HBV/ F isolates into two subtypes (F1 and F2) would be more appropriate than providing them with two distinct genotypes (F and H). 展开更多
关键词 GENOTYPES Hepatitis B virus PHYLOGENETICANALYSIS SUBTYPES
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Screening of hepatocyte proteins binding to F protein of hepatitis C virus by yeast two-hybrid system 被引量:2
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作者 Yan-Ping Huang Jun Cheng +10 位作者 Shu-Lin Zhang Lin Wang Jiang Guo Yan Liu Yuan Yang Li-Ying Zhang Gui-Qin Bai Xue-Song Gao Dong ji Shu-Mei Lin Qing Shao 《World Journal of Gastroenterology》 SCIE CAS CSCD 2005年第36期5659-5665,共7页
AIM: To investigate the biological function of F protein by yeast two-hybrid system. METHODS: We constructed F protein bait plasmid by cloning the gene of F protein into pGBKT7, then recombinant plasmid DNA was tran... AIM: To investigate the biological function of F protein by yeast two-hybrid system. METHODS: We constructed F protein bait plasmid by cloning the gene of F protein into pGBKT7, then recombinant plasmid DNA was transformed into yeast AH109 (a type). The transformed yeast AH109 was mated with yeast Y187 (α type) containing liver cDNA library plasmid in 2×YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-HisAde) containing X-α-gal for selection and screening. After extracting and sequencing plasmids from positive (blue) colonies, we underwent sequence analysis by bioinformatics. RESULTS: Thirty-six colonies were selected and sequenced. Among them, 11 colonies were zymogen granule protein, 5 colonies were zinc finger protein, 4 colonies were zinc-α-2-glycoprotein, 1 colony was sialyltransferase, 1 colony was complement control protein factor I, 1 colony was vitronectin, and 2 colonies were new genes with unknown function. CONCLUSION: The yeast two-hybrid system is an effective method for identifying hepatocyte proteins interacting with F protein of hepatitis C virus. F protein may bind to different proteins. 2005 The WJG Press and Elsevier Inc. All rights reserved 展开更多
关键词 Hepatitis C virus f protein Yeast two-hybridsystem
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