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81.
DING Xuedong WANG Guohua PAN Xiangchen LIU Dongdong LUO Xuedong MA Yuan LI Maolin CUI Qi ZHANG Qijin 《中国畜牧兽医》2007,47(12):3815-3824
The purpose of this study was to investigate the variation of Orf virus (ORFV) immune related genes after infection with different species.The ORFV genomes of sheep and camel were extracted and named ORFV-Y and ORFV-LT,respectively.Based on ORFV genome sequence published in GenBank (accession No.:KF234407.1),three pairs of specific primers were designed and synthesized to amplify the B2L,F1L and VIR gene fragments of ORFV-Y and ORFV-LT,respectively,and the amplified fragments were cloned into pMD19-T vector,transformed into E.coli DH5α competent cells.The recombinant plasmid was identified,and positive clones were selected for sequencing,DNAStar software was used to analyze the homology,amino acid sequence and phylogenetic tree of 13 ORFV genome sequences published on NCBI.The results showed that the nucleotide homology of B2L,F1L and VIR genes were 92.8% to 99.2%,95.7% to 99.5% and 77.6% to 100%,respectively.After comparing the amino acid sequence between the two genomes and the reference sequence,it was found that there were obvious differences in the immune related genes between the two genomes,and F1L gene had some rules to follow.The phylogenetic analysis of B2L,F1L and VIR genes showed that ORFV-Y was closely related to the Chinese Fujian goat strain,while ORFV-LT was far from the reference strains,and was a separate branch.The results showed that ORFV had obvious difference in immune related genes between sheep and camels,it provided a reference basis for further research on the changes of ORFV gene sequences in different species and the development of vaccines for different species in the future. 相似文献
82.
This study was aimed to analyze the genetic variation of insulin-like growth factor binding protein 2 (IGFBP2) gene effect on meat quality traits in pigs. A pair of primer was designed,which based on DNA sequence of IGFBP2 gene (GenBank accession No.:BV727778). The genetic polymorphism of IGFBP2 gene was detected by PCR-RFLP in Sujiang pig, Jiangquhai pig and Landrace pig, and the association between IGFBP2 gene polymorphism and meat quality traits in three pig breeds was analyzed. The results showed that three genotypes (AA, AB and BB) were detected. The Chi-square test results showed that the genotype distribution of three pig breeds was in Hardy-Weinberg equilibrium (P > 0.05). The polymorphism information content (PIC) were moderately polymorphic in three pig breeds. According to the association analysis of different genotypes, significant difference was found in color value, intramuscular fat content and marbling, the color value (except for Jianquhai pig) and intramuscular fat content of AA genotype were significantly higher than AB and BB genotypes (P < 0.05), the marbling of AA and AB genotypes was significantly higher than BB genotype (P < 0.05). In conclusion,the polymorphism of IGFBP2 gene intron 3 had significant effects on meat quality traits, and was candidate gene for the potential impact on meat quality traits of pigs. 相似文献
83.
84.
Theo W. Prins Lia Wagemakers Jan A.L. van Kan 《European journal of plant pathology / European Foundation for Plant Pathology》2000,106(7):693-698
To identify genes of the necrotrophic pathogenic fungus Botrytis
cinerea that are expressed during infection of tomato leaves, a differential screening of a genomic library with radioactively labelled cDNA was performed. This resulted in the identification of a B. cinerea gene, denominated Bcubi4, which encodes a precursor protein consisting of four identical head-to-tail repeats of a 76aa ubiquitin unit. Subsequently a gene denominated Bcubi1CEP79, encoding a single ubiquitin unit joined to a Carboxyl Extension Protein of 79 amino acids, was isolated. The expression of the two ubiquitin genes was studied during pathogenesis of B. cinerea on tomato. Bcubi1CEP79, but not Bcubi4, mRNA was transiently induced at 16h after inoculation. The increased expression of the Bcubi1CEP79 gene at this stage of pathogenesis might be required for enhanced ribosomal biogenesis. 相似文献
85.
沃尔巴克氏体(Wolbachia)主要感染昆虫等节肢动物,是一类革兰阴性共生细菌。为探讨昆虫纲共生菌Wolbachia的系统发育,研究Wolbachia基因组中进化速度较快的外膜蛋白基因wsp的密码子使用模式及影响因素,统计分析该基因的GC含量(GCall、GC1、GC2、GC3、GC3s)、有效密码子数(ENc)、相对同义密码子使用度(RSCU)等指标。结果显示:昆虫纲共生菌Wolbachia的wsp基因密码子偏好性均不强,昆虫纲不同目间共生菌Wolbachia的wsp基因碱基组成及ENc值差异较小;多数基因数据点沿标准曲线或在其附近分布,突变对碱基组成的影响较弱;18种由多个密码子编码的氨基酸中有11种氨基酸的偏好性密码子在昆虫纲7个目及对照蛛形纲1个目间均相同,有2种氨基酸的偏好性密码子在7个目间相同,这些密码子均以A/U结尾;供分析的140个wsp基因中仅编码6个半胱氨酸(Cys);对应分析中第1、第2向量轴贡献率均不高,分别为13.53%和12.49%,均与碱基组成(GC1、GC2、GC3)显著相关。综合各项分析认为,Wolbachia的wsp基因密码子偏好性不具有宿主分类特异性,密码子使用模式主要受碱基组成影响,而碱基组成主要受选择影响。 相似文献
86.
为了解广西地区猪繁殖与呼吸综合征病毒(PRRSV)流行毒株的遗传进化情况,对2014年-2016年来自广西各地的部分PRRSV阳性病料进行Nsp2和ORF5基因的扩增和测序分析.结果获得34个Nsp2基因序列和45个ORF5基因序列,均属于美洲型毒株.Nsp2基因间核苷酸序列的同源性为91.8%~100%,与PRRSV美洲型毒株VR-2332、CH-1a、JXA1及NADC30株核苷酸序列的同源性分别为81.3%~84.3%、88.9%~92.1%、94.3%~99.3%和73.5%~75.1%,而与PRRSV欧洲型毒株LV株核苷酸序列的同源性为51.5%~53.2%.ORF5基因间核苷酸序列的同源性为82.8%~100%,与PRRSV美洲型毒株VR-2332、CH-1a、JXA1及NADC30株核苷酸序列的同源性分别为83.7%~99.5%、85%~95%、83.8%~99.7%和83.2%~86.4%,而与PRRSV欧洲型毒株LV株核苷酸序列的同源性为62.4%~64.5%.基于Nsp2和ORF5基因推导的氨基酸序列绘制的遗传进化树中,广西地区的毒株主要分布在以JXA1为代表的Ⅳ亚群.表明当前广西PRRSV流行毒株以JXA1株为代表的高致病性美洲型毒株为主,各毒株Nsp2和ORF5基因序列存在一定的差异,尚未发现欧洲型毒株和美洲型NADC30类毒株. 相似文献
87.
88.
香蕉枯萎病是由尖孢镰刀菌古巴专化型(Fusarium oxysporum f. sp. cubense,Foc)引起的香蕉毁灭性土传病害,其中 4号生理小种(Foc4)能感染几乎所有的香蕉品系,危害最严重。SMART在线软件分析myosin-1基因具有肌球蛋白马达蛋白(myosin motor domain, MMD),肌动蛋白尾结构TH1(myosin tail)和 Src家族同源结构域SH3(src homology domain 3),与禾谷镰刀菌中氰烯菌酯靶标基因myosin-5具高度的蛋白同源性,相似性高达83%。利用Split-marker基因重组技术获得Foc4的myosin-1基因敲除突变体,Δmyosin-1突变株丧失了对氰烯菌酯的敏感性,菌丝生长缓慢,产孢量减少且孢子畸形,对香蕉致病力严重下降,证实myosin-1是氰烯菌酯在Foc4中的作用靶标基因。外施靶向myosin-1体外转录的dsRNA,能抑制菌丝的生长,降低菌丝活性;菌丝膨胀扭曲分枝增多,出现典型的球状结构,与氰烯菌酯处理后的表型一致。在盆栽活体人工接种实验中,体外施用dsRNA可以明显抑制枯萎病外部症状的发展,推迟发病时间,赋予寄主抗性,结果说明体外施用dsRNA可以作为新型杀菌剂防治香蕉枯萎病。综上,myosin-1基因作为氰烯菌酯在Foc4中的靶标基因具有高度的序列保守,在调控菌丝生长发育,产孢以及致病力等方面发挥重要作用,而外施dsRNA具有防治香蕉枯萎菌的巨大潜力。 相似文献
89.
PuPR为含有编码人孕激素受体(hPR)激素结合区(LBD)基因(631-933氨基酸)的pUC19重建质拉,在其宿主Escherichiacoli(BL21)中,经1mmol/L的IPTG诱导,该转化的细菌产生一C端为6×His结尾的蛋白质,分子量45KD。该表达产物可利用Ni-NTAHRP在ng水平用Western印迹检测,并可通过亲和层析的方法,利用His-bindingresin(Ni树脂)从菌体的裂解液中方便地回收到该蛋白质。 相似文献
90.
苹果柱型基因Co的一个AFLP 标记的SCAR转换 总被引:15,自引:4,他引:15
将苹果柱型基因的一个AFLP 标记成功地转换成了简单实用的SCAR 标记。首先对AFLP 标记片段进行序列测定, 然后根据序列特点设计了两对特异引物CoA1/ CoA2 和CoA1/ CoA3 , 每条引物长20 bp。PCR 结果表明CoA1/ CoA2 可以扩增出216 bp 和148 bp 两条带, 其中216 bp 的为柱型性状的特征带; CoA1/CoA3 可以扩增出273 bp 和205 bp 的两条带, 其中273 bp 的为柱型性状的特征带。两对引物在杂交后代中扩增出的特征带与柱型性状的分离重组率都很低(CoA1/ CoA2 为6. 3 % ±2. 5 %; CoA1/ CoA3 为7. 3 % ±2. 6 %) , 所以它们都可以作为该SCAR 标记的特异引物所用。 相似文献