排序方式: 共有26条查询结果,搜索用时 15 毫秒
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纳豆激酶基因在毕赤酵母中的表达纯化及抗体制备 总被引:3,自引:0,他引:3
目的实现纳豆激酶(NK)在巴斯德毕赤酵母中的分泌表达,以纯化产物为抗原制备兔抗NK血清。为建立双抗夹心酶联免疫吸附反应法测定生物体内NK含量,进一步研究NK在体内代谢与功能奠定基础。方法将NK基因克隆到毕赤酵母表达载体pHBM905A中,鉴定后的重组质粒用SalⅠ酶切线性化后电转化到毕赤酵母宿主菌GS115中,甲醇诱导表达,经盐析和Millipore超滤膜过滤分离纯化表达产物。纤维蛋白法检测纯化产物的活性,并以纯化产物为抗原免疫新西兰大白兔制备兔抗NK血清。结果SDS-PAGE结果显示NK成功表达,其相对分子质量(Mr)约为27 000,纤维蛋白平板实验显示表达产物具有较好的纤溶活性。酶联免疫法测得多克隆抗体效价约为1∶8 000,Western blot结果显示在Mr27 000附近有1条特异带出现。结论NK在巴斯德毕赤酵母中实现了分泌表达,表达产物具有较好的纤溶活性和免疫原性。 相似文献
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纳豆激酶粗提液对家兔实验性高脂血症的作用研究 总被引:1,自引:0,他引:1
目的:研究纳豆激酶粗提液对家兔实验性高脂血症的调节作用。方法:用猪油和胆固醇喂饲家兔造成高脂血症模型,造模同时分A、B组分别添加120mL/d和120mL/d纳豆激酶粗提液,8周后采静脉血测定血清总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C),计算动脉粥样硬化指数AI。结果:实验中高脂饮食可成功诱导高脂血症(P0.01)。纳豆激酶粗提液高剂量组(B组)TC、TG、LDL-C值均显著低于模型组(P0.05),HDL-C值极显著高于模型组(P0.01),与纳豆激酶粗提液低剂量组(120Ml/d)相比,高剂量组(240mL/d)降低TC、TG值效果显著(P0.05)。结论:纳豆激酶粗提液对脂质代谢失调有明显的改善和调节作用。 相似文献
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Zijing Huang Tsz Kin Ng Weiqi Chen Xiaowei Sun Dingguo Huang Dezhi Zheng Jingsheng Yi Yanxuan Xu Xi Zhuang Shaolang Chen 《Investigative ophthalmology & visual science》2021,62(6)
PurposeNattokinase (NK), an active ingredient extracted from traditional food Natto, has been studied for prevention and treatment of cardiovascular diseases due to various vasoprotective effects, including fibrinolytic, antihypertensive, anti-atherosclerotic, antiplatelet, and anti-inflammatory activities. Here, we reported an antineovascular effect of NK against experimental retinal neovascularization.MethodsThe inhibitory effect of NK against retinal neovascularization was evaluated using an oxygen-induced retinopathy murine model. Expressions of Nrf2/HO-1 signaling and glial activation in the NK-treated retinae were measured. We also investigated cell proliferation and migration of human umbilical vein endothelial cells (HUVECs) after NK administration.ResultsNK treatment significantly attenuated retinal neovascularization in the OIR retinae. Consistently, NK suppressed VEGF-induced cell proliferation and migration in a concentration-dependent manner in cultured vascular endothelial cells. NK ameliorated ischemic retinopathy partially via activating Nrf2/HO-1. In addition, NK orchestrated reactive gliosis and promoted microglial activation toward a reparative phenotype in ischemic retina. Treatment of NK exhibited no cell toxicity or anti-angiogenic effects in the normal retina.ConclusionsOur results revealed the anti-angiogenic effect of NK against retinal neovascularization via modulating Nrf2/HO-1, glial activation and neuroinflammation, suggesting a promising alternative treatment strategy for retinal neovascularization. 相似文献
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目的:通过对纳豆发酵中加入中药苦参、板蓝根、漏芦、红花后纳豆激酶活力的改变进行研究,从而筛选出能够提高纳豆激酶活力的中药,同时也将中药之药性融入其中,以便更好地提高纳豆之医疗、保健功能。方法:本实验采用纤维蛋白-琼脂糖平板法测定发酵后产物中纳豆激酶的活力大小。结果:苦参、板蓝根可提高纳豆激酶活性,漏芦可降低纳豆激酶活性,红花可抑制枯草芽孢杆菌的生长使之没有活性。结论:发酵纳豆中加入中药后对纳豆激酶的活力有不同程度的影响。 相似文献
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纳豆激酶基因重组质粒在大肠杆菌与毕赤酵母中的稳定性 总被引:10,自引:0,他引:10
利用PCR方法以纳豆杆菌染色体DNA为模板扩增纳豆激酶基因片段,分别克隆到原核表达载体pBV220与真核表达截体pPICZaA上,转化大肠杆菌HB101与毕赤酵母GS115,筛选重组子,通过限制性内切酶,PCR分析及序列测定,测定该重组子所携外源基因为纳豆激酶基因,对重组大肠杆菌与毕赤酵母中外源基因的稳定性进行研究比较,结果表明外源基因的插入对宿主菌的生长没有太大影响,重组质粒在E.coli HB101中具有良好的分离稳定性,而结构稳定性较差,而外源基因在毕赤酵母(GS115中很稳定。 相似文献
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《Yao wu shi pin fen xi = Journal of food and drug analysis.》2014,22(4):485-491
In this study we have evaluated the changes in arterial blood pressure in spontaneously hypertensive rats (SHR) caused by the short-term intake of Bacillus subtilis B060-fermented beans with significant γ-aminobutyric acid (GABA) and nattokinase activity. After being weaned, 7-week-old male SHR and 7-week-old male Wistar–Kyoto (WKY) rats were randomized into seven groups. Until the 8th week of life, the rats in each group were given one of the following: Group 1, high dose of GABA and nattokinase in the SHR (SHD); Group 2, medium dose of GABA and nattokinase in the SHR (SMD); Group 3, low dose of GABA and nattokinase in the SHR (SLD); Group 4, negative control in the SHR (SD); Group 5, positive control in the SHR (SM); Group 6, high dose of GABA and nattokinase in the WKY (WHD); and Group 7, negative control in the WKY (WD). Distilled water served as the negative control, and captopril (50 mg/kg), a known ACE inhibitor, served as the positive control. Systolic blood pressure and diastolic blood pressure values were measured weekly from the 8th week to the 16th week of life using the tail-cuff method. A definite decrease in systolic and diastolic blood pressure values could be observed in the rats treated with captopril and in the rats that received GABA and nattokinase. The greatest antihypertensive effect was observed when the pharmacological treatment was administered. The effect of the daily intake of fermented beans containing GABA and nattokinase may be helpful in controlling blood pressure levels in hypertensive model animals. The fermentation of beans with B. subtilis B060 may therefore constitute a successful strategy for producing a functional food with antihypertensive activity. 相似文献
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