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AIM: To investigate the effect of transketolase-like protein 1 (TKTL1) on proliferation of human nasopharyngeal carcinoma cells in vitro. METHODS: The siRNA against TKTL1 mRNA was constructed and transfected into human nasopharyngeal carcinoma cells (CNE cell line). The activity of transketolase was detected before and after RNA interference.Real-time PCR was used to determine the mRNA expression of transketolase (TKT) gene family in the CNE cells.Flow cytometry and MTT test were used to detect the effect of anti-TKTL1 siRNA on cell proliferation and cell cycle in the CNE cells. RESULTS: The total transketolase activity was significantly decreased in the CNE cells transfected with siRNA TKTL1 construct compared with the cells transfected with control vector or untransfected CNE cells. No significant difference in the expression level of TKT and TKTL2 gene between the CNE cells transfected with siRNA TKTL1 construct and the cells transfected with control vector or untransfected CNE cells was observed (P>0.05). However, the expression level of TKTL1 gene was significantly downregulated in the CNE cells transfected with siRNA TKTL1 construct compared with the cells transfected with control vector.Cancer cells were arrested in G0/G1 phase, and cancer cell proliferation was significantly inhibited in the CNE cells transfected with siRNA TKTL1 construct. CONCLUSION: TKTL1 plays an important role in the total transketolase activity and cell proliferation of human nasopharyngeal carcinoma. TKTL1 may be considered as a potential target for novel anti-cancer therapy. 相似文献
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嗜水气单胞菌J-1株OmpA融合蛋白的高效表达及免疫原性分析 总被引:3,自引:0,他引:3
以嗜水气单胞菌(Aeromonas hydrophila)J-1株基因组为模板,根据GenBank已发表的嗜水气单胞菌外膜蛋白的基因序列设计1对引物,扩增去除信号肽序列的成熟外膜蛋白OmpA的基因片段,定向克隆至表达质粒pET32a(+)中,重组质粒经限制性酶切鉴定后测序。结果表明,插入片段长度为948bp,与NCBI上登录的几个相关序列相比,同源性在93.1%-95%之间,缺失4个氨基酸。将重组原核表达质粒pET32a-OmpA转化至大肠杆菌BL21(DE3),经诱导可表达分子量约57.4kD的蛋白,凝胶薄层扫描显示OmpA在转化菌中表达量占全菌蛋白的50%以上。用兔抗J-1株总外膜蛋白血清进行Western blot,在57.4kD处有明显反应条带。融合蛋白经Ni-NTA树脂柱亲和层析纯化后免疫新西兰兔制备抗血清,ELISA效价达1:12800以上。Western blot检测结果显示,该血清与9株具有代表性的气单胞菌的分子量约35kD的外膜蛋白均有较强反应,说明所表达的融合蛋白不仅保持原有外膜蛋白的免疫原性,而且提示OmpA可能是嗜水气单胞菌的共同保护性抗原。[中国水产科学,2008,15(2):301—306] 相似文献
226.
Effects of different dietary protein and lipid levels on growth, feed utilization and body composition of red porgy (Pagrus pagrus) fingerlings 总被引:1,自引:0,他引:1
D. SCHUCHARDT J.M. VERGARA H. FERNÁNDEZ-PALACIOS C.T. KALINOWSKI C.M. HERNÁNDEZ-CRUZ M.S. IZQUIERDO & L. ROBAINA 《Aquaculture Nutrition》2008,14(1):1-9
Two feeding trials were conducted to determine the minimum dietary protein level producing maximum growth, and the optimum protein to energy ratio in diets for red porgy (Pagrus pagrus) fingerlings, respectively. In the first trial, six isoenergetic diets were formulated with protein levels ranging from 400 to 650 g kg?1 in increments of 50 g kg?1, and fed for 11 weeks to 2.8 g average initial weight fish. Weight gain, specific growth rate and feed efficiency were significantly higher with diets containing higher protein levels, when compared with dietary levels below 500 g kg?1. The highest protein efficiency ratios were obtained in fish fed 500 g kg?1 dietary protein. The minimum dietary protein level producing maximum fish growth was found to be 500 g kg?1. In the second trial, 15 g average initial weight fish were fed for 12 weeks, six diets containing three different lipid levels (100, 150 and 200 g kg?1) combined with two protein levels (450 and 500 g kg?1). Weight gain values increased when dietary lipids increased from 100 to 150 g kg?1, with a further decrease for 200 g kg?1 lipids in diets; the lowest fish growth being supported by 200 g kg?1 dietary lipids. Fish growth was significantly higher when dietary protein increased from 450 to 500 g kg?1. There was no evidence of a protein‐sparing effect of dietary lipids. Liver protein and lipid contents were low when compared with other fish species. All diet assayed produced high liver glycogen accumulation. The recommended protein and lipid levels in diets for red porgy fingerlings were 500 and 150 g kg?1, respectively. 相似文献
227.
A 10‐week feeding experiment was conducted to evaluate the effect of different protein to energy ratios on growth and body composition of juvenile Litopenaeus vannamei (initial average weight of 0.09 ± 0.002 g, mean ± SE). Twelve practical test diets were formulated to contain four protein levels (300, 340, 380 and 420 g kg?1) and three lipid levels (50, 75 and 100 g kg?1). Each diet was randomly fed to triplicate groups of 30 shrimps per tank (260 L). The water temperature was 28.5 ± 2 °C and the salinity was 28 ± 1 g L?1 during the experimental period. The results showed that the growth was significantly (P < 0.05) affected by dietary treatments. Shrimps fed the diets containing 300 g kg?1 protein showed the poorest growth. However, shrimp fed the 75 g kg?1 lipid diets had only slightly higher growth than that fed 50 g kg?1 lipid diets at the same dietary protein level, and even a little decline in growth with the further increase of dietary lipid to 100 g kg?1. Shrimp fed the diet with 420 g kg?1protein and 75 g kg?1 lipid had the highest specific growth rate. However, shrimp fed the diet with 340 g kg?1 protein and 75 g kg?1 lipid showed comparable growth, and had the highest protein efficiency ratio, energy retention and feed efficiency ratio among dietary treatments. Triglycerides and total cholesterol in the serum of shrimp increased with increasing dietary lipid level at the same dietary protein level. Body lipid and energy increased with increasing dietary lipid level irrespective of dietary protein. Results of the present study showed that the diet containing 340 g kg?1 protein and 75 g kg?1 lipid with digestible protein/digestible energy of 21.1 mg kJ?1 is optimum for L. vannamei, and the increase of dietary lipid level has not efficient protein‐sparing effect. 相似文献
228.
C. FERNANDES da SILVA E. BALLESTER J. MONSERRAT L. GERACITANO W. WASIELESKY JR P.C. ABREU 《Aquaculture Nutrition》2008,14(6):507-514
The nutritional quality of biofilm, a microbial community associated to an organic matrix, was evaluated in artificial substrate (polyethylene screen) in net cages during 30 days in the Patos Lagoon estuary, Southern Brazil. During this period, samples of biofilm were collected each 5 days for analysis of chlorophyll a, microorganisms abundance, dry weight, protein and lipid contents. During the study, chlorophyll a varied from 0.38 to 2.75 μg cm?2; dry weight between 7.16 and 17.63 mg cm?2; protein content from 0.43 to 1.76 mg cm?2 and lipid concentration between 1.21 and 4.23 mg cm?2. The variation of lipid in the biofilm was closely related to the abundance of free heterotrophic bacteria (34.25–56.54 × 106 cells cm?2), filamentous cyanobacteria (7.5–15.9 × 106 filaments cm?2), flagellates (6.92–12.89 × 106 cells cm?2) and mainly nematodes (29–1,414 organisms cm?2), while protein content varied similarly to the abundance of unicellular centric diatoms (52.10–179.81 × 103 cells cm?2), and nematodes. This information will allow a better management of food supply to raised aquatic organism with the utilization of natural productivity in the culture systems, with considerable decrease in production costs. 相似文献
229.
通过生理性解偶联调节氧化磷酸化效率可能是进化早期发展起来的一种普遍策略,对生命的存在至关重要. 解偶联蛋白(uncoupling protein,UCP)家族是位于线粒体内膜的转运蛋白,哺乳动物基因组中已发现5个成员,其中UCP1~UCP3紧密相关,而UCP4和UCP5(即BMCP1,brain mitochondrial carrier protein-1)与它们差异较大. UCP1~UCP4不仅在哺乳类、鸟类等恒温脊椎动物中普遍存在,而且在鱼类等变温动物中也被证实存在,但UCP5目前仅在哺乳类中被发现. 哺乳动物褐色脂肪组织的UCP1,介导呼吸链产生的质子梯度泄漏,使氧化产生的能量以热的形式散发,但鱼类等变温动物UCP1的功能尚待进一步研究确立. UCP2和UCP3在产热、控制活性氧生成、脂肪酸氧化以及肥胖、糖尿病的发生中均发挥重要作用,二者的转录调节较复杂. UCP4和UCP5与其他UCP关系较远,UCP4仅在脑中表达,而UCP5在脑中高表达,它们的功能还不十分明确,但研究表明很可能与UCP2和UCP3的功能相似. 相似文献
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