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将抗癌胚抗原(CEA)单克隆抗体的重链可变区与人的恒定区(Cγ3)连接,制备抗癌胚抗原嵌合重链用于放免治疗及其他导向治疗,可减少人抗鼠抗体反应(HAMA)。为纯化及核素标记抗体,将嵌合重链基因与核心链霉亲和素基因融合。融合基因在大肠杆菌得到高效表达,表达量占菌体总蛋白的24%。SDS-PAGE和蛋白质印迹图谱显示表达产物分子量为70kD,与其基因编码蛋白质的理论推算值相符。以HRP标记的生物素作为 相似文献
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将抗癌胚抗原单链抗体基因与核心链霉亲和素基因融合插入昆虫杆状病毒供体质粒pFastBacHTa中,在粉纹夜蛾Tn-5B1-4细胞中进行表达。SDS-PAGE分析结果表明,表达产物分子量为41kD左右,Western印迹分析结果表明,以HRP标记的生物素进行蛋白质印迹在41kD处可见表达条带,表明融合蛋白能特异性的与生物素结合,放射免疫分析表明重组杆状病毒表达产生的ScFv-CS蛋白能特异性结合癌胚 相似文献
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抗癌胚抗原单链抗体基因的构建及高效表达 总被引:8,自引:2,他引:6
采用重组PCR技术将已获得的抗癌胚抗原鼠源单克隆抗体E7B10重,轻链可变区基因经(Gly3Ser)3编码的寡核苷酸拼接成单链抗体基因,并在大肠杆菌中高效表达了单链抗体C端的融合蛋白,其表达量达到菌体总蛋白的20%,SDS-PAGE和Western印迹图谱显示表达产物分子量为27kD,与其基因编码蛋白的理论推算值相符。 相似文献
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将抗癌胚抗原单链抗体基因与核心链霉亲和素基因融合插入昆虫杆状病毒供体质粒 pFastBacHTa中 ,在粉纹夜蛾Tn 5B1 4细胞中进行表达。SDS PAGE分析结果表明 ,表达产物分子量为 4 1kD左右 ,Western印迹分析结果表明 ,以HRP标记的生物素进行蛋白质印迹在 4 1kD处可见表达条带 ,表明融合蛋白能特异性的与生物素结合 ,放射免疫分析表明重组杆状病毒表达产生的ScFv CS蛋白能特异性结合癌胚抗原 相似文献
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本文采用反转录-聚合酶链反应(RT-PCR)技术,从鼠抗乙肝病毒表面抗原(HBsAg)单克隆细胞中克隆到了该抗体重、轻链可变区(Ⅴ区)基因,并分别将其与人的恒定区基因Cγ3,Ck相拼接,构建人-鼠嵌合抗体基因。SDS-PAGE和Western-Blot分析结果证实嵌合抗体重链基因在E.coli中得到了表达。间接ELISA法免疫测定的结果表明该表达产物具有与乙肝表面抗原结合的能力。 相似文献
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乙肝表面抗原专一的单链嵌合抗体在大肠杆菌中的表达 总被引:3,自引:0,他引:3
利用重组PCR技术将乙肝表面抗原专一单抗的Vk与人源的C基因拼接成轻链嵌合抗体V-C,再通过编码柔性肽段(Gly4Ser)的Linker与V连接成单链嵌合抗体ScFV-C,并分别在大肠杆菌的热敏与分泌型表达系统中进行表达。表达产物的Western-blot与间接ELISA检测结果表明,SCFv-C产两种系统中的表达产物均具有抗原结合活性,并已在分泌肽的指导下ScFv-Ck能被E.coli分泌出胞外。 相似文献
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组织型纤溶酶原激活剂A链与尿激酶原B链嵌合分子的构建与表达 总被引:1,自引:0,他引:1
利用 D N A 重组技术和定位删除技术,将组织型纤溶酶原激活剂(t P A)的 A 链( Serl Thr263)基因与尿激酶原(pro U K)的 B链( Ser138 Leu411)基因相连,得到嵌合分子基因tu pa,并在昆虫杆状病毒系统中进行表达,表达量可达 500 I U/m l.经单克隆抗体免疫亲和层析纯化细胞表达上清液,得到tu P A 嵌合分子,其比活为 200 000 I U/m g 蛋白. S D S P A G E 及 W estern blot 鉴定证明此表达产物分子量约为 60 k D,与预期值相符.纤维蛋白平板测活及纤维蛋白亲和性分析初步证明,此嵌合分子的溶纤活性与pro U K 相近,而纤维蛋白亲和性高于 pro U K. 相似文献
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将抗癌胚抗原(CEA)单链抗体基因插入家蚕杆状病毒转移载体pBacPAKHis, 与修饰的家蚕核型多角体病毒BmBacPAKDNA共转染家蚕细胞, 经同源重组得到含有在多角体蛋白基因启动子控制下的抗CEAScFv 基因的重组病毒BmBacScFv。用重组病毒分别感染家蚕细胞和幼虫, 在两者中均得到了高效表达, 产物分子量为28kD, 前者占细胞总蛋白的6 % , 后者为0 .3 mg/ 蚕。目的基因在家蚕细胞和幼虫中表达产物经Ni2+IDASepharose6B亲和柱纯化, 前者纯度可达90% 以上, 后者纯度较低; 纯化后的融合蛋白具有CEA 结合活力, 其亲和常数分别为5 .4×108/mol·L- 1 和2.3 ×108/mol·L-1 , 略低于其亲本单抗E7B10 2.7 ×109/mol·L- 1 。 相似文献
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牛肾上腺皮质LDL受体经Triton X-100增溶,DEAE32离子交换柱和LpB Sepharose亲和柱层析,在SDS-PAGE中有三条区带,分别在原点;Mr 160kD;Mr125kD处。进一步用8%SDS-PAGE纯化三个区带的蛋白质分别免疫新西兰大白兔所得的抗体,应用免疫印迹和ECL非同位素标记法可对牛肾上腺皮质和人皮肤纤维细胞膜上的LDL受体进行测定。 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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Michael Hesse 《Plant Systematics and Evolution》1980,134(3-4):229-267
Some closely related members of the monocotyledonous familiesAlismataceae, Liliaceae, Juncaceae, Cyperaceae, Poaceae andAraceae with variable modes of pollination (insect- and wind-pollination) were studied in relation to the ultrastructure of pollenkitt and exine (amount, consistency and distribution of pollenkitt on the surface of pollen grains). The character syndromes of pollen cementing in entomophilous, anemophilous and intermediate (ambophilous or amphiphilous) monocotyledons are the same in principal as in dicotyledons. Comparing present with former results one can summarize: 1) The pollenkitt is always produced in the same manner by the anther tapetum in all angiosperm sub-classes. 2) The variable stickiness of entomophilous and anemophilous pollen always depends on the particular distribution and consistency of the pollenkitt, but not its amount on the pollen surface. 3) The mostly dry and powdery pollen of anemophilous plants always contains a variable amount of inactive pollenkitt in its exine cavities. 4) A step-by step change of the pollen cementing syndrome can be observed from entomophily towards anemophily. 5) From the omnipresence of pollenkitt in all wind-pollinated angiosperms studied one can conclude that the ancestors of anemophilous angiosperms probably have been zoophilous (i.e. entomophilous) throughout. 相似文献
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Surveillance of Class Ⅰ Newcastle Disease Virus at Live Bird Markets and Commercial Poultry Farms in Eastern China Reveals the Epidemic Characteristics 下载免费PDF全文
Xiaolong Lu Xiaoquan Wang Tiansong Zhan Yifan Sun Xin Wang Naiqing Xu Tianxing Liao Yu Chen Min Gu Shunlin Hu Xiaowen Liu Xiufan Liu 《中国病毒学》2021,36(4):818-822
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正Dear Editor,Parainfluenza virus 5 (PIV5), known as canine parainfluenza virus in the veterinary field, is a negative-sense,nonsegmented, single-stranded RNA virus belonging to the Paramyxoviridae family (Chen 2018). The virus was first reported in primary monkey kidney cells in 1954 (Hsiung1972), then it has been frequently discovered in various 相似文献
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Altaf Hussain Tiantian Wu Hui Li Linjin Fan Kai Li Li Gao Yongqiang Wang Yulong Gao Changjun Liu Hongyu Cui Qing Pan Yanping Zhang Asim Aslam Khan Muti-Ur-Rehman Muhammad Munir Salman Latif Butt Xiaomei Wang Xiaole Qi 《中国病毒学》2019,34(1):102-105
<正>Dear Editor,Infectious bursal disease (IBD) is one of the most important diseases of the poultry. The IBD virus (IBDV), a nonenveloped virus belonging to the Birnaviridae family with a genome consisting of two segments of double-stranded RNA (segments A and B), targets B lymphocytes of bursa of Fabricious leading to immunosuppression. In Pakistan,poultry farming is the second biggest industry and IBD is the second biggest disease threating the poultry sector.However, there is limited genome information of IBDV 相似文献