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1.
本研究旨在探明鸡恒定链(invariant chain,Ii)与内吞体转运蛋白Rab5a和Rab7b结合的结构域和在细胞内共定位的特征。首先,用PCR和基因突变技术将Ii胞浆区与跨膜区[Ii(Cyt-Tra)]、Ii CLIP (class Ⅱ-associated invariant chain peptide)-三聚体区[Ii(CLIP-TRIM)]和Ii突变体[Ii(M81-87aa)、Ii(M91-99aa)和Ii(M81-99aa)]分别插入pET-32a和pEGFP-C1构建相应的原核和真核重组质粒。其次,将构建的含有绿色荧光蛋白的重组质粒与实验室保存的含有红色荧光Rab5a和Rab7b的重组质粒共转染至人胚胎肾细胞系293 T,观察它们的共定位。将构建的原核重组质粒进行表达和纯化,最后用拉下法和免疫印迹检测Ii与Rab5a和Rab7b的结合域。结果表明,成功构建Ii结构域及Ii突变体的重组质粒。Ii(Cyt-Tra)及Ii突变体均能与Rab5a和Rab7b在细胞内共定位,而Ii(CLIP-TRIM)与空载体却不能。Ii的胞浆区和跨膜区是与Rab5a和Rab7b结合的功能结构域,而不是CLIP与三聚体区。综上所述,鸡Ii与Rab5a和Rab7b共定位和结合的区域是其胞浆区和跨膜区,而不是内质网腔区。这些结果提示Rab分子参与了Ii在胞内细胞器的转运机制,为进一步研究Ii及其载体在细胞内的转运机制和功能提供了新的途径。 相似文献
2.
分泌抗鸡Ig单抗杂交瘤细胞系的建立与单抗生物学特性的鉴定 总被引:7,自引:0,他引:7
从传染性法氏囊病病毒(IBDV)单克隆抗体(McAb)杂交瘤细胞系1D10、1G1中得到了3株能稳定分泌抗鸡免疫球蛋白(Ig)McAb的杂交瘤细胞系1B7、1D7、3G6。其抗体类和亚类均属IgG2b,腹水的ELISA效价≥1∶25600,琼扩效价为1∶8~1∶16。3株McAb能与鸡血清中的Ig出现沉淀反应,琼扩在24h以内出现清晰的沉淀线,而不能和鸭、鸽、鹌鹑等禽类及异种动物血清出现沉淀线。纯化的鸡IgG、IgM经SDS-PAGE后,分别用纯化并经辣根过氧化物酶(HPR)标记的1B7、1D7、3G6单抗酶结合物进行免疫印迹试验证明,3株单抗识别的均为IgG、IgM分子的轻链 相似文献
3.
研究了催化氯化的产物分布,试验了有关条件对产物分布的影响。结果表明:氯化程度是产物分布的主要影响因素;温度的提高,有利于苄叉二氯和苄川三氯的生成;铁离子、低温及水会导致环上的氯化;水引起水解的发生。 相似文献
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ZHOU Shu-lu YE Ren-gao LIU Xiao-bo ZHANG Hong XU Han-shi DU Yong LI You-ji YANG Nian-sheng YANG Xiao YU Xue-qing 《园艺学报》2003,19(6):782-785
AIM:To detect the association between the polymorphism of Fc receptor γ chain gene at position-29 in promoter and systemic lupus erythematosus(SLE).METHODS:The genotypes at position -29 in promoter of Fc receptor γ chain gene were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method in 180 patients with SLE and 140 ethnically matched controls in southern China.RESULTS:The frequencies of TT genotype(33.3%) and T allele (54.4%) at position -29 in patients with SLE were significantly higher than those in controls (17.2% and 42.9%, respectively), whereas, the frequencies of GG genotype (24.4%) and G allele (45.6%) in patients with SLE were remarkably lower than those in controls (31.4% and 57.1%, respectively) (P<0.05). The TT genotype and T allele at position -29 were not associated with lupus nephritis in SLE patients (P>0.05).CONCLUSION:Our results indicate that the T allele at position -29 in promoter of Fc receptor gene probably contributes to the susceptibility to SLE, but does not play a role in the occurrence of lupus nephritis. 相似文献
6.
P. Galeffi G. Giunta S. Guida C. Cantale 《European journal of plant pathology / European Foundation for Plant Pathology》2002,108(5):479-483
Citrus tristeza virus (CTV) is one of the most destructive citrus virus diseases in the world. The construction of an engineered antibody, EMBL accession number AJ278109, able to specifically recognize its antigen, i.e. the coat protein of CTV, directly on infected plant material without any purification or manipulation of the entire woody plant. The potential uses of this engineered antibody are discussed. 相似文献
7.
Seasonal distribution of phytoplasmas in Australian grapevines 总被引:1,自引:0,他引:1
The distribution and persistence of phytoplasmas were determined in Australian grapevines. Phytoplasmas could be detected using the polymerase chain reaction (PCR) from shoots, cordons, trunks and roots throughout the year, and phytoplasmas appear to persistently infect Australian grapevines from year to year. Phytoplasmas were not always detected in samples from the same sampling area from one sampling period to the next. Phytoplasma detection by PCR was improved by sampling from shoots, cordons and trunks, especially during October (early spring). The diseases expressed by the 20 grapevines used in the distribution and persistence studies were monitored. Australian grapevine yellows disease (AGY) was expressed by 17/20 grapevines at some time during the study, whilst only 4/20 and 15/20 grapevines expressed restricted growth disease (RG) and late season leaf curl disease (LSLC), respectively. All grapevines with RG and LSLC also had AGY. The three diseases were persistently expressed in some grapevines and remission of disease was observed in others. The results of PCR detection in the same grapevines indicated that phytoplasmas were more frequently detected in AGY-affected grapevines that also expressed RG and LSLC compared with grapevines expressing AGY alone. Phytoplasmas were detected in symptomless plant material but less frequently compared with AGY-affected material. 相似文献
8.
Mariner转座子与小菜蛾抗性关系的研究 总被引:1,自引:1,他引:1
以抗溴氰菊酯、杀虫双、杀螟丹小菜蛾种群及其敏感品系为研究对象,借助聚合酶链式反应(PCR)、载体筛选、琼脂糖凝胶电泳等实验技术,检测了小菜蛾4个品系的Mariner转座子的存在及其与抗性的关系。结果表明,在抗溴氰菊酯、杀虫双、杀螟丹以及敏感品系的小菜蛾中都存在两种大约500 bp的Mariner转座子片断,初次证明在小菜蛾4个品系中均含有两种Mariner转座子,并发现一种新的转座子基因片断,但是没有发现Mariner转座子与抗性之间存在直接的联系。研究结果将为利用Mariner转座子标签法在小菜蛾中分离定位基因、转化外源基因、转基因昆虫防治害虫等研究提供理论基础。 相似文献
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10.
Branched‐chain amino acid ratios in low‐protein diets regulate the free amino acid profile and the expression of hepatic fatty acid metabolism‐related genes in growing pigs 下载免费PDF全文
Y. H. Duan F. N. Li C. Y. Wen W. L. Wang Q. P. Guo Y. H. Li Y. L. Yin 《Journal of animal physiology and animal nutrition》2018,102(1):e43-e51
Liver metabolism is affected by nutrients. The aim of this study was to explore the effects of low‐protein diets (17% crude protein, CP) supplemented with branched‐chain amino acids (BCAAs), including leucine (Leu), isoleucine (Ile) and valine (Val), on hepatic amino acid profile and lipid metabolism in growing pigs. The ratio of Leu : Ile : Val in all groups was 1 : 0.51 : 0.63 (20% crude protein, CP), 1 : 1 : 1 (17% CP), 1 : 0.75 : 0.75 (17% CP), 1 : 0.51 : 0.63 (17% CP) and 1 : 0.25 : 0.25 (17% CP) respectively. Results revealed that compared to the positive control group (1 : 0.51 : 0.63, 20% CP), the low‐protein diets significantly augmented the concentrations of most essential amino acids and non‐essential amino acids (p < .05), with the greatest values observed in the 1 : 0.25 : 0.25 group. Moreover, relative to the control, the low‐protein diets with the Leu : Ile : Val ratio ranging from 1 : 0.75 : 0.75 to 1 : 0.25 : 0.25 markedly downregulated the mRNA abundance of acetyl‐CoA carboxylase (ACC), lipoprotein lipase (LPL) and fatty acid‐binding protein 4 (FABP‐4) (p < .05), and upregulated the mRNA expression of hormone‐sensitive lipase (HSL), peroxisome proliferator‐activated receptor‐g coactivator‐1α (PGC‐1α), uncoupling protein 3 (UCP3) and liver carnitine palmitoyltransferase 1 (L‐CPT‐1) (p < .05). Therefore, our data suggest that protein‐restricted diets supplemented with optimal BCAA ratio, that is, 1 : 0.75 : 0.75–1 : 0.25 : 0.25, induce a shift from fatty acid synthesis to fatty acid oxidation in the liver of growing pigs. These effects may be associated with increased mitochondrial biogenesis. 相似文献