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马白细胞介素18成熟蛋白(mEIL-18)基因在大肠埃希氏菌中的高效表达与纯化 总被引:2,自引:0,他引:2
用RT—PCR从经ConA刺激的马外周血单个核细胞(PBMC)中扩增出马白细胞介素18(Equine interleukin-18,EIL-18)前体蛋白(precursor EIL-18,pEIL-18)基因的cDNA,然后克隆至载体pCR2.1-TOPO中,鉴定并命名为pCR2.1-pEIL-18。自pCR2.1-pEIL-18中扩增马白细胞介素18成熟蛋白(mature EIL-18,mEIL-18)基因,并将其亚克隆至原核表达载体pET-28a(+)中。将筛选出的阳性克隆进行测序、诱导表达并纯化其表达产物。结果表明,mEIL-18基因全长474bp,含1个开放阅读框,编码157个氨基酸的成熟蛋白;表达产物以可溶性和包涵体两种形式存在,经SDS-PAGE和Western—blot分析,重组蛋白相对分子量约为20ku,且具有免疫生物学活性。mEIL-18基因在大肠杆菌中高效表达并在非变性和变性条件下采用Ni^+亲和层析纯化方法获得了高纯度的重组mEIL-18,为探究马白细胞介素18的生物学活性及其应用奠定了基础。 相似文献
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为研究肉牛对不同玉米秸杆产品的利用情况,选18月龄西门塔尔牛与当地黄牛的F136头,分别用铡碎、揉碎、揉碎压捆、制粒、压块和挤压成棒状的六种玉米秸秆饲料进行了肉牛饲养试验和消化试验.结果秸秆采食量以后三种加工处理的秸秆为最高,铡碎为最低,揉碎和揉碎压捆居中;日粮表现消化率以秸秆颗粒型日粮最低,其他几种秸秆型日粮差异不大;肉牛增重以应用秸秆饲料块和秸秆饲料棒的肉牛增重效果最好,铡碎秸秆最差,揉碎和揉碎压捆居中.除铡碎型外,其他几种玉米秸杆饲料用于育肥牛均可取得较好的增重效果. 相似文献
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对四个香菇开放式接种场所内不同时间点环境中霉菌和细菌的数量、温度和湿度进行测定,结果表明,在同一测试场所,不同测试时间点环境中霉菌和细菌的数量有所变化。总体上,消毒前和接种结束时环境中霉菌和细菌的数量相对较多,接种开始和接种进行一半时环境中霉菌和细菌的数量较少。各场所环境中的温度、湿度与霉菌和细菌的数量无相关性。消毒前环境中霉菌、细菌数量与菌棒污染率成正相关,相关系数分别为0.83和0.96;接种开始时环境中霉菌、细菌数量与菌棒污染率的相关系数分别为0.64和0.94;接种进行一半和接种结束时环境中的霉菌数量与菌棒污染率无相关性,而这两个时间点细菌数量与菌棒污染率的相关系数分别为0.62和0.88。 相似文献
87.
WENG Wen-jun FANG Jian-pei WU Yan-feng XIAO Lu-lu XU Hong-gui BAO Rong WEI Jing XU Lü-hong 《园艺学报》2007,23(11):2200-2204
AIM: To probe the effect of different panel reactive antibody(PRA) serum levels from patients with β-thalassemia on the proliferation and differentiation potential of the hematopoietic stem /progenitor cell of cord blood.METHODS: 1×105 mononuclear cells (MNCs) isolated from umbilical cord blood were incubated with different PRA serum levels (0 μL,50 μL,100 μL) respectively and complement,inoculated into the methylcellulose cultural system.The proliferation and differentiation potential of the hematopoietic stem /progenitor cell of cord blood by the colony formation assay were detected on day 7 and day 14,respectively.RESULTS: After culture of 7 days,the total colonies and CFU-GM were 88.20±9.41,79.00±11.39 in group A and 88.60±9.12,79.20±10.44 in group B,which were significantly higher than those of 20.60±7.39,15.20±4.66 in group C and those of 4.00±2.05,1.40±0.51 in group D (P<0.01).Meanwhile compared with group A and group E,no significant difference was found (P>0.05).After culture for 14 days,the total colonies and CFU-GM were 216.00±31.10,117.40±24.80 in group A and 213.20±31.06,116.00±19.75 in group B,which were significantly higher than those of 97.80±14.43,32.80±8.10 in group C and those of 31.40±13.41,8.40±4.30 in group D (P<0.01).The CFU-GEMMs were 45.60±8.51 in group A and 42.60±7.03 in group B,which were significantly higher than those of 20.80±6.96 in group C and those of 7.80±6.06 in group D (P<0.05).The BFU-MK was 12.80±4.42 in group A and 11.00±2.74 in group B respectively,which were significantly higher than that of 1.00±0.55 in group D (P<0.05).The CFU-E in group B 17.20±4.03 was significantly higher than that of 5.60±2.87 in group D (P<0.05).Meanwhile compared with group A and group E,no significant difference was found (P>0.05).By the Kendall test,there were negative correlations between the level of PRA serum and the total colonies,CFU-GM on day 7,the total colonies,CFU-GM,CFU-GEMM,BFU-E,BFU-MK on day 14 (tau-b=-0.793,-0.849,-0.808,-0.804,-0.645,-0.674,-0.624,P<0.01).There was a negative correlation between the level of PRA serum and CFU-MK on day 14 (tau-b=-0.466,P<0.05).CONCLUSION: PRA sera inhibit the colony in the colony cultures of the hematopoietic stem cells/progenitor cells in cord blood.The inhibition depends on the level of PRA sera.The higher the level of PRA sera,the stronger the inhibition is observed in our study. 相似文献
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分析了黄山市生态旅游的资源优势,探讨了生态旅游资源开发中所存在的问题,从法制建设、科学规划、人才建设等方面提出了相关的建议及对策。 相似文献
90.
Effects of fruit bagging on coloring and related physiology,and qualities of red Chinese sand pears during fruit maturation 总被引:1,自引:0,他引:1
Chunhui Huang Bo Yu Yuanwen Teng Jun Su Qun Shu Zaiquan Cheng Liqiong Zeng 《Scientia Horticulturae》2009
Red Chinese sand pears (Pyrus pyrifolia Nakai) are particular to China. In order to determine the effects of fruit bagging treatments (including bag types, bag removal patterns and dates) on fruit qualities and to understand the mechanism of coloring of red Chinese sand pears, two experiments were carried out. In the first experiment, fruit of ‘Meirensu’ were firstly covered by light-impermeable paper bags with different levels of light permeable liners during their early development stage, then, the whole bag were/not removed or only the outer layer of bags were removed 3, 2 or 1 weeks before harvest. Thus, the fruit were/not totally re-exposed or were under different levels of sunlight transmission (80.31% or 34.71%). Non-bagged fruit were used as the control. Bagging treatments significantly affected the concentration of anthocyanin and the visual qualities of pear fruit. Compared to control, fruit re-exposed totally for 2 or 3 weeks accumulated the largest amount of anthocyanin and fruit receiving 80.31% and 34.71% of sunlight for 1–3 weeks could synthesize a little anthocyanin, indicating that high light intensity is imperative for coloring in red Chinese sand pears. Bagging treatments did not affect contents of total soluble sugars, but decreased organic acids contents in fruit. In the second experiment, fruit of ‘Meirensu’ and ‘Yunhongli No. 1’ were covered with only one type of light-impermeable bag during the early development stage and totally re-exposed after the bag removal 15 days before harvest. Fruit were then collected at different intervals to trace the time-course of coloring, and related physiology and inner qualities. With increasing time after the bag removal, the concentration of chlorophyll, carotenoid, flavonoid and total phenols changed little, but the concentration of anthocyanin accumulated extremely fast within 10 days after the bag removal in both cultivars and thereafter kept constant. ‘Yunhongli No. 1’ had higher anthocyanin contents and lower hue angle than ‘Meirensu’, indicating a higher potential of anthocyanin synthesis. After the bag removal, the sucrose contents and PAL activities increased gradually and correlation analysis revealed that they were highly correlated with anthocyanin accumulation in two cultivars. This study suggests that anthocyanin biosynthesis in red Chinese sand pears is a highly light dependent process and modified by genotypes. Based on the current results, in order to obtain red Chinese sand pear fruit with attractive appearance and good inner qualities, fruit must be covered with light-impermeable bags at the early stage of fruit development and the bag should be removed totally at least 10 days before harvest. 相似文献