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大肠杆菌表达的重组人粒细胞-巨噬细胞集落刺激因子的纯化 总被引:1,自引:0,他引:1
对重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)高效表达克隆pZW.GM的表达产物进行了纯化,并对纯化的GM-CSF进行了N端氨基酸序列分析。人GM-CSF基因表达产物在大肠杆菌中以不溶性包涵体形式存在,经过超声破菌、包涵体抽提、凝胶过滤层析、复性、离子交换一系列化步骤,终产物纯度达99%,按蛋白总量计算回收率达10%,比活性达1×10^7u/mg蛋白质。通过测定纯化人GM-CSF的N端1 相似文献
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人PSP基因在昆虫细胞中的高效表达 总被引:2,自引:0,他引:2
应用昆虫杆状病毒表达系统(BES)在昆早细胞Tn-5B1-4中高效表达了人persephin(PSP),SDS-PAGE分析表达量占细胞可深性蛋白质的20%左右,表达产物经Ni^2+-NTA树脂亲和层析纯化后纯度达85%以上。活性研究表明,昆早细胞表达的PSP蛋白能显著促进脊髓神经元的存活。 相似文献
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应用疏水层析对大肠杆菌表达的人重组白细胞介素-4(rhIL-4)进行了纯化,含有rhIL-4的包涵体,经洗涤、变性、复性后,以Butyl-Sepharose层析,得到了高纯度的rhIL-4.纯度达97%;回收率为32%;比活性为2×10~7U/mg,讨论了rhIL-4疏水层析的条件,并对不同的方法纯化白细胞介素-4进行了比较. 相似文献
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以0.1mol/LNH4Cl溶液为介质,用2.5次微分伏安法测定了丙二醛,线性范围为1.0*10^-6至1.0*10^-3mol/L,检测限达1.0*10^-7mol/L。并测定了细胞培养液介质中新生SD大鼠室肌细胞样品的丙二醛。 相似文献
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建立了一个实验室制备重组hGM-CSF的培养、复性和分离纯化的技术工艺。通过对PEGM工程菌的生长和诱导表达等条件的初步调整,使hGM-CSF的表达量从16.9%提高到32.3%,获得0.749g/L的包含体,纯度为65%;包含体用6mol/L盐酸胍溶解和稀释复性后,经CMSepharoseFF.DEAESepharoseFF和SephacrylS200HR层析分离,制得纯度达97%以上、比活性为3.0×107u/mg的hGM-CSF,纯化总回收率约为5% 相似文献
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用羊β-乳球蛋白基因(BLG)5’区5*10^3b(10^3b,旧称kb)为调序列,构建了乳腺表达组织纤溶酶原激活剂突变体载体。对540枚小鼠受精卵进行显微注射,经PCR和Southernblot检测,获得6只整合有人La-tPA的转基因小鼠,整合率为32%。 相似文献
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采用高密度发酵法发酵重组人γ干扰素产生菌SWINF/DH5α并成功地获得高表达、高产量的菌体,表达的γ干扰素蛋白占菌体总蛋白的60%以上,20L发酵液可收获2kg(湿重)菌体。纯化过程中采用尿素抽提、稀释、复性、浓缩后通过SephacrylS200柱的纯化方法,简化了纯化步骤,缩短周期,提高了蛋白回收率。纯化中未采用单抗亲和层析技术,使产品更为安全可靠。最终产品经SDSPAGE检测纯度达100%,核酸含量和热原均合格,A280/A260>15,比活性达1×107IU/mg,总回收率达40%。这说明整个工艺适宜大规模生产的需要,具有实际应用价值。 相似文献
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亲和层析纯化HepG2细胞分泌的PAI—1 总被引:1,自引:0,他引:1
本文报道用抗PAI-1单克隆抗体(McAb)亲和层析建立了纯化PAI-1的简便方法。经免疫亲和层析,Sephacryl S200凝胶过滤,从HepG2细胞培液中分离到糖基化和非糖基化两种形式的PAI-1,回收率为84%,PAI-1比活性为6.1×10^4IU/mg。糖基化PAI-1分子量为50kD,比活性5.8×10^4IU/mg。非糖基化PAI-1分子量43kD,占总PAI-130%,仍具有PA 相似文献
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Curcumin is the yellow pigment of turmeric that interacts irreversibly forming an adduct with thioredoxin reductase (TrxR), an enzyme
responsible for redox control of cell and defence against oxidative stress. Docking at both the active sites of TrxR was performed to compare
the potency of three naturally occurring curcuminoids, namely curcumin, demethoxy curcumin and bis-demethoxy curcumin. Results show
that active sites of TrxR occur at the junction of E and F chains. Volume and area of both cavities is predicted. It has been concluded by
distance mapping of the most active conformations that Se atom of catalytic residue SeCYS498, is at a distance of 3.56 from C13 of
demethoxy curcumin at the E chain active site, whereas C13 carbon atom forms adduct with Se atom of SeCys 498. We report that at least
one methoxy group in curcuminoids is necessary for interation with catalytic residues of thioredoxin. Pharmacophore of both active sites of
the TrxR receptor for curcumin and demethoxy curcumin molecules has been drawn and proposed for design and synthesis of most probable
potent antiproliferative synthetic drugs. 相似文献
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正Dear Editor,In December 2019, a novel human coronavirus caused an epidemic of severe pneumonia(Coronavirus Disease 2019,COVID-19) in Wuhan, Hubei, China(Wu et al. 2020; Zhu et al. 2020). So far, this virus has spread to all areas of China and even to other countries. The epidemic has caused 67,102 confirmed infections with 1526 fatal cases 相似文献
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The young pistils in the melanthioid tribes, Hewardieae, Petrosavieae and Tricyrteae, are uniformly tricarpellate and syncarpous. They lack raphide idioblasts. All are multiovulate, with bitegmic ovules. The Petrosavieae are marked by the presence of septal glands and incomplete syncarpy. Tepals and stamens adhere to the ovary in the Hewardieae and the Petrosavieae but not in the Tricyrteae. Two vascular bundles occur in the stamens of the Hewartlieae and Tricyrtis latifolia. Ventral bundles in the upper part of the ovary of the Hewardieae are continuous with compound septal bundles and placental bundles in the lower part. Putative ventral bundles occur in the alternate position in the Tricyrteae and putative placental bundles in the opposite. position in the Petrosavieae. The dichtomously branched stigma in each carpel of the Tricyrteae is supplied by a bifurcated dorsal bundle. 相似文献
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鸡传染性法氏囊病病毒研究进展 总被引:3,自引:0,他引:3
传染性法氏囊病(Infection bursal disease, IBD)是由鸡传染性法氏囊病毒(Infectious bursal disease virus, IBDV)引起的鸡和火鸡的一种高度接触性传染病,给世界各国的禽养殖业带来了巨大损失.自IBDV发现至今新的变异株不断出现,分子结构的改变导致病毒致病力的改变及宿主对疫苗应答的改变,使得传统的疫苗已不能控制其流行,因此各国学者对其基因组结构和功能进行了广泛深入的研究,并积极研制新型有效的疫苗以达到防治的目的. 相似文献
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Renfei Lu Xiuming Wu Zhenzhou Wan Yingxue Li Lulu Zuo Jianru Qin Xia Jin Chiyu Zhang 《中国病毒学》2020,35(3):344-347
In conclusion, the novel visual RT-LAMP assay is a simple, rapid, and sensitive approach for detection of SARS-CoV-2, and it is ready for application in primary care and community hospitals or health care centers, and even patients' own houses in response to the current SARS-CoV-2 epidemic because the assay does not require sophisticated equipment and skilled personnel. Furthermore, it is also ready to be used in fields for screening samples from wild animals and environments to facilitate the identification of potential intermediate hosts that mediate the cross-species transmission of SARS-CoV-2 from bats to humans. 相似文献
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Shen Jia-Yuan Li Man Xie Lyu Mao Jia-Rong Zhou Hong-Ning Wang Pei-Gang Jiang Jin-Yong An Jing 《中国病毒学》2021,36(1):145-148
正Dear Editor,Chikungunya virus (CHIKV), an arbovirus in the family of Togaviridae, genus Alphavirus, is transmitted by the A.aegyptii or A. albopictus mosquito, and causes disease in humans characterized by fever, rash, and arthralgia (Silva and Dermody 2017; Suhrbier 2019). It was first reported in 1953 in Tanzania, and caused only a few outbreaks and sporadic cases in Africa and Asia in last century. However, in the epidemic in 2004, CHIKV acquired mutations that conferred enhanced transmission by the A. albopictus mosquito(Schuffenecker et al. 2006). Since then, it has successively caused outbreaks in Africa, the Indian Ocean, South East Asia, the South America, and Europe (Zeller et al. 2016). 相似文献