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鸡白细胞介素-6原核表达及双抗夹心ELISA方法的建立 总被引:1,自引:0,他引:1
应用分子生物学技术原核表达ChIL-6,以ChIL-6重组蛋白为免疫原,按免疫程序分别制备兔抗和鼠抗IL-6重组蛋白的多克隆抗体。应用此抗体建立双抗夹心EL-ISA方法后,为了优化此方法本试验对该方法的最佳试验条件、标准曲线、重复性和初步应用进行确定。结果显示,经SDS-PAGE和Western-blot分析,表明ChIL-6在大肠杆菌中正确表达,为了建立检测ChIL6的双抗夹心ELISA方法,本试验应用表达的重组蛋白制备兔抗和鼠抗多克隆抗体。建立的双抗夹心ELISA方法最佳反应条件为包被抗体的质量浓度为50mg/L,4℃过夜;酶标二抗稀释度为1:400,37。C1h;应用该方法检测感染金黄色葡萄球菌后的ChIL-6,其结果与本实验室之前应用人的ELISA试剂盒检测的结果相似。结果表明,本试验建立的检测ChlL-6的双抗夹心ELISA方法可用于临床。 相似文献
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YU Si-yang WANG Yan ZENG Gao-feng LIU Yang XU Jian-qiang ZENG Meng-ya TANG Ye-hua ZENG Zhi-ying SHI Xiao-qiao CHEN Ying ZHAO Guo-jun 《园艺学报》2016,32(5):863-868
AIM: To explore the role of nucleotide-binding oligomerization domain-like receptor protein 1 (NLRP1) inflammasome in atorvastatin-induced reduction of interleukin-1β (IL-1β) and interleukin-18 (IL-18) releases from the THP-1 macrophages. METHODS: Lipopolysaccharide (LPS, 10 μg/L) was used to trigger the secretion of IL-1β and IL-18 in the THP-1 macrophages. The cells were incubated with different concentrations of atorvastatin (1, 10 and 20 μmol/L) for 24 h, or treated with 10 μmol/L atorvastatin for different time (12 h, 24 h and 48 h). NLRP1 siRNA was transfected into the THP-1 cells. The mRNA expression of NLRP1 inflammasome was detected by RT-PCR. The protein expression of NLRP1 inflammasome was determined by Western blot. The secretion of proinflammatory cytokines IL-1β and IL-18 was quantified by ELISA. RESULTS: Atorvastatin inhibited the mRNA and protein expression of NLRP1 inflammasome in the THP-1 macrophages in a dose- and time-dependent manner. Transfection of NLRP1 siRNA significantly decreased the protein expression of NLRP1 and promoted the suppressive effect of atorvastatin on IL-1β and IL-18 secretion in the THP-1 macrophages. CONCLUSION: Atorvastatin inhibits the production of IL-1β and IL-18 in the macrophages through decreasing NLRP1 inflammasome expression, possibly contributing to the anti-inflammatory effect of atorvastatin on atherosclerosis. 相似文献
34.
辣椒新品种航椒18号的选育 总被引:2,自引:0,他引:2
航椒18号是以051-3-2-2-H-H-H为母本,以035-1-2-2-H-H为父本杂交育成的早中熟羊角型辣椒一代杂种。从定植至青熟果采收55d(天)左右,始花节位为第9~11节。果面皱、果肩宽、有光泽,味香辣;单株结果数20个左右,单果质量50.5g左右;果实粗长羊角形,纵径22.7cm,横径4.2cm,肉厚0.25cm。田间对辣椒病毒病、炭疽病和疫病的抗性强于对照陇椒3号。平均每667m~2产量4200kg,适宜在甘肃、内蒙古、新疆等地及生态环境相似区域保护地栽培。 相似文献
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根据已经发表的F18ab菌毛A亚单位(FedA/ab)的基因(fedA/ab)[1],设计一对引物,利用PCR技术从表达F18ac菌毛的大肠杆菌2134P株[2]、8199株[3]、8813株[3]中分别扩增到一段序列,并克隆至pGEM-T载体,获得重组质粒T2134PA、T8199A、T8813A.琼脂糖凝胶电泳、序列测定及分析表明,该3个序列大小均为516bp,与fedA/ab(513bp)具有较高的同源性,分别为96.3%、96.5%、95.9%,推导的Fed/ac氨基酸序列与FedA/ab同源性分别为93.0%、93.6%、92.4%.数据表明该实验所克隆的序列均为F18ac菌毛A亚单位(FedA/ac)的基因(fedA/ac). 相似文献
37.
OBJECTIVE: Genetic and biological characterisation of 12 isolates of Cryptosporidium from pigs and comparing them with Cryptosporidium isolates from humans and cattle. DESIGN: Cryptosporidium isolates from pigs were compared with those obtained from human and cattle using rDNA sequence analysis. The infectivity of two of the porcine isolates was determined in neonatal mice and the clinical history of the infected pigs recorded. RESULTS: Pig-derived isolates of Cryptosporidium exhibited two distinct genotypes; a porcine genotype and a bovine genotype, which is common to cattle and other livestock. The porcine genotype did not produce any infection in neonatal mice whereas the bovine genotype did. CONCLUSION: Two distinct genetically and biologically differing strains of Cryptosporidium appeared to be associated with acute diarrhoea in pigs. Whether Cryptosporidium was a primary or secondary pathogen is unclear but warrants further investigation. As the bovine genotype is known to infect humans, the results suggest that pigs can act as reservoirs of cryptosporidial infections for humans and other live-stock. The zoonotic potential of the pig-adapted genotype is uncertain and requires further study. 相似文献
38.
Martin-Laurent F Piutti S Hallet S Wagschal I Philippot L Catroux G Soulas G 《Pest management science》2003,59(3):259-268
We report the development of quantitative competitive (QC) PCR assays for quantifying the 16S, 18S ribosomal and atzC genes in nucleic acids directly extracted from soil. QC-PCR assays were standardised, calibrated and evaluated with an experimental study aiming to evaluate the impact of atrazine application on soil microflora. Comparison of QC-PCR 16S and 18S results with those of soil microbial biomass showed that, following atrazine application, the microbial biomass was not affected and that the amount of 16S rDNA gene representing 'bacteria' increased transitorily, while the amount of 18S rDNA gene representing fungi decreased in soil. In addition, comparison of atzC QC-PCR results with those of atrazine mineralisation revealed that, in response to atrazine treatment, the amount of atzC gene increased transitorily in soil pre-treated with atrazine, suggesting that accelerated atrazine biodegradation in soil could be due to a transient increase in the size of the atrazine mineralising community. 相似文献
39.
ELIZABETH A. BALLEGEER LISA J. FORREST ROBERT JERAJ T. ROCKWELL MACKIE R. JEROME NICKLES 《Veterinary radiology & ultrasound》2006,47(2):228-233
A primary lung tumor in a dog treated with intensity-modulated radiation therapy was imaged approximately 6 weeks and 1-year posttreatment with combined positron emission tomography (PET) and computed tomography, utilizing the radiotracers 18F-fluorodeoxyglucose and 18F-fluorothymidine. These two tracers allowed discrimination of tumor from inflammation, and demonstrated spread of tumor along airways over time after treatment. Fusion of functional imaging with anatomic imaging is a useful tool, particularly in the field of oncology, with the potential for PET markers that delineate tumor from normal or reactive tissue, and potential or actual response to therapy. 相似文献
40.