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表达流感病毒神经氨酸酶基因的重组腺病毒的构建 总被引:1,自引:0,他引:1
通过RT-PCR方法扩增流感病毒神经氨酸酶基因,将其克隆到腺病毒穿梭载体pTrackCMV,此重组质粒与腺病毒DNA共转化大肠杆菌BJ5183,通过细菌内同源重组获得重组腺病毒DNA,将其转染293细胞获得重组腺病毒.经PCR证实目的基因已整合至腺病毒基因组中,western blot检测到神经氨酸酶的表达.重组病毒经滴鼻和灌胃两种途径免疫小鼠,结果表明2次免疫后滴鼻组和灌胃组均产生明显的免疫应答反应,滴鼻组的免疫效果优于灌胃组. 相似文献
104.
Feng YL 《Physiologia plantarum》2008,132(3):350-358
There is a trade-off between nitrogen (N) allocation to photosynthesis and to defence. Invasive species may reduce N allocation to defence because of the absence of natural enemies. Thus, I hypothesised that invasive species may allocate a higher fraction of total leaf N to photosynthesis and have higher light-saturated photosynthetic rate ( P max ) and photosynthetic N-use efficiency (PNUE) than closely related native species. To test these hypotheses, invasive Eupatorium adenophorum and native E. chinense and E. heterophyllum were compared in a limestone shrub. Unlike expectation, the invader did not allocate a higher fraction of leaf N to photosynthesis than the natives. However, it was more efficient in photosynthetic N partitioning than the natives. It partitioned a higher fraction of the photosynthetic N to carboxylation and showed higher use efficiency of the photosynthetic N, while the natives partitioned a higher fraction of the photosynthetic N to light-harvesting components. Total leaf N content was not significantly different among the three studied invasive and native species. For the invader, the higher fraction of leaf N allocated to carboxylation resulted in the higher N content in carboxylation and in both carboxylation and bioenergetics, which led to higher P max , and therefore to higher PNUE, water-use efficiency, respiration efficiency and apparent quantum yield. These physiological advantages of the invader and its higher leaf area ratio may contribute to its invasiveness. 相似文献
105.
Jia XY Guo ZY Wang Y Xu Y Duan SS Feng YM 《Protein science : a publication of the Protein Society》2003,12(11):2412-2419
The single-chain insulin (PIP) can spontaneously fold into native structure through preferred kinetic intermediates. During refolding, pairing of the first disulfide A20-B19 is highly specific, whereas pairing of the second disulfide is likely random because two two-disulfide intermediates have been trapped. To get more details of pairing property of the second disulfide, four model peptides of possible folding intermediates with two disulfides were prepared by protein engineering, and their properties were analyzed. The four model peptides were named [A20-B19, A7-B7]PIP, [A20-B19, A6-B7]PIP, [A20-B19, A6-A11]PIP, and [A20-B19, A7-A11]PIP according to their remaining disulfides. The four model peptides all adopt partially folded structure with moderate conformational differences. In redox buffer, the disulfides of the model peptides are more easily reduced than those of the wild-type PIP. During in vitro refolding, the reduced model peptides share similar relative folding rates but different folding yields: The refolding efficiency of the reduced [A20-B19, A7-A11]PIP is about threefold lower than that of the other three peptides. The present results indicate that the folding intermediates corresponding to the present model peptides all adopt partially folded conformation, and can be formed during PIP refolding, but the chance of forming the intermediate with disulfide [A20-B19, A7-A11] is much lower than that of forming the other three intermediates. 相似文献
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Yabin Wang Kazim Narsinh Li Zhao Dongdong Sun Dongjuan Wang Zheng Zhang Zhongchan Sun Rongqing Zhang Haichang Wang Feng Cao 《Cell biology international》2011,35(2):135-140
Ghrelin is thought to directly exert a protective effect on the cardiovascular system, specifically by promoting vascular endothelial cell function. Our study demonstrates the ability of ghrelin to promote rat CMEC (cardiac microvascular endothelial cell) proliferation, migration and NO (nitric oxide) secretion. CMECs were isolated from left ventricle of adult male Sprague—Dawley rat by enzyme digestion and maintained in endothelial cell medium. Dil‐ac‐LDL (1,1′‐dioctadecyl‐3,3,3′,3′‐ tetramethylindocarbocyanine‐labelled acetylated low‐density lipoprotein) intake assays were used to identify CMECs. Cells were split into five groups and treated with varying concentrations of ghrelin as follows: one control non‐treated group; three ghrelin dosage groups (1×10?9, 1×10?8, 1×10?7 mol/l) and one ghrelin+PI3K inhibitor group (1×10?7 mol/l ghrelin+20 μmol/l LY294002). After 24 h treatment, cell proliferation capability was measured by MTT [3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐2H‐tetrazolium bromide] assay and Western blot for PCNA (proliferating cell nuclear antigen) protein expression. Migration of CMECs was detected by transwell assays, and NO secretion of CMECs was measured via nitrate reduction. Protein expression of AKT and phosphorylated AKT in CMECs was measured by Western blot after exposure to various concentrations of ghrelin and the PI3K inhibitor LY294002. Our results indicate that ghrelin significantly enhanced cell growth at concentrations of 10?8 mol/l (0.271±0.041 compared with 0.199±0.021, P=0.03) and 10?7 mol/l (0.296±0.039 compared with 0.199±0.021, P<0.01). However, addition of the PI3K/AKT inhibitor LY294002 inhibited the ghrelin‐mediated enhancement in cell proliferation (0.227±0.042 compared with 0.199±0.021, P=0.15). At a concentration between 10?8 and 10?7 mol/l, ghrelin caused a significant increase in the number of migrated cells compared with the control group (126±9 compared with 98±7, P=0.02; 142±6 compared with 98±7, P<0.01), whereas no such change could be observed in the presence of 20 μmol/l of the PI3K/Akt inhibitor LY294002 (103±7 compared with 98±7, P=0.32). Ghrelin treatment significantly enhanced NO production in a dose‐dependent fashion compared with the untreated control group [(39.93±2.12) μmol/l compared with (30.27±2.71) μmol/l, P=0.02; (56.80±1.98) μmol/l compared with (30.27±2.71) μmol/l, P<0.01]. However, pretreatment with 20 μmol/l LY294002 inhibited the ghrelin‐stimulated increase in NO secretion [(28.97±1.64) μmol/l compared with (30.27±2.71) μmol/l, P=0.37]. In summary, we have found that ghrelin treatment promotes the proliferation, migration and NO secretion of CMECs through activation of PI3K/AKT signalling pathway. 相似文献
110.
以植物钾离子外排通道(K’channeloutward.rectifier,KCO)基因为研究对象,运用CodonW软件分析了75个植物KCO基因密码子的使用模式,探讨密码子的使用模式和影响密码子使用的各种可能因素。结果表明:碱基组成差异(r=0.961,P〈0.01)和自然选择(r=0.568,P〈0.01)是影响密码子使用的主要因素,并且高表达的基因强烈偏爱使用以G或C结尾的密码子。确定了UUC、CUC等26个均以G/C结尾的密码子为植物KcD基因的高表达优越密码子。 相似文献