Cloning, Expression and Polyclonal Antibody Preparation of the Asialoglycoprotein Receptor of Marmot

来源 :华中科技大学学报(医学)(英德文版) | 被引量 : 0次 | 上传用户:wpsx236
下载到本地 , 更方便阅读
声明 : 本文档内容版权归属内容提供方 , 如果您对本文有版权争议 , 可与客服联系进行内容授权或下架
论文部分内容阅读
The objective of this study is to express the carbohydrate recognition domain (CRD) of the asialoglycoprotein receptor (ASGPR) H1 and H2 subunits of Marmota himalayan in vitro, and develop polyclonal antibodies against the recombinant proteins. RT-PCR was used to amplify ASGPR CRDH1 and CRDH2 from the liver tissue of Marmota himalayan. The products of amplification were subcloned into prokaryotic expression vector pRSET-B, and expressed in E. coli BL21(DE3)plysS. The recombinant proteins were purified using Ni-NTA spin column. The purified proteins were inoculated into BALB/c mice to develop polyclonal antibodies. The sensitivity and specificity of antibodies were evaluated by enzyme-linked immunosorbent assay (ELISA), Western blotting and immunohistochemical staining (IHC). The polyclonal antibodies showed high sensitivity and specificity against both denaturated and native ASGPR proteins. We successfully amplified and expressed the ASGPR CRDs of Marmota himalayan. The nucleic sequences of ASGPR CRDH1 and CRDH2 of Marmota himalayan have been submitted to Genbank and the sequence ID are DQ 845465 and DQ845466, respectively. The proteins and antibodies prepared can be used for targeting gene therapy in a new animal model-Marmota himalayan-for the research of infectious diseases of hepatitis viruses and liver cancer treatment.
其他文献
每当黄昏时分,我做完作业便会搬来小凳子坐在院子里,开始欣赏夕阳那迷人的景色。起初还蓝蓝的天空,这时已变成了淡黄色。也只是持续了一会儿,金黄色的晚霞就气势恢宏地登场了
Submerged arc welding (SAW) has shortcoming of over heat input, which increases grain size in heat-affected zone (HAZ) and decreases welding quality and mechani
在古代诗文的教学中,用人性论的观点去考察人性的境界,我以为对于一个教师来说,是很有必要的。因为,在教学中对学生人格的发展将产生重大影响。就我理解的人性境界来说,我以
近日,由国内多家文学网站推选的“中国十大‘80后’作家主编”榜单出炉,郭敬明、韩寒、张悦然等“写而优则编”的文坛80后旗帜人物纷纷入榜,然而,独占榜首的却是一位来自陕西
《木兰诗》是我国古典诗歌中的一首脍炙人口的优秀诗篇。它以浓厚真挚的感情和铺叙、对照渲染的表现手法,集中描绘了一个传奇的女子——木兰代父从军的故事。文中塑造了一个
新华每日电讯等多家媒体报道,在成都被誉为“粽子一条街”的马鞍东路上,有商家挂出一句“雷人”的广告词,“屈原大叔别跳了,吃个胖姐粽子吧”。网友质疑这广告有不尊重屈原之
近日,作家余华、导演孟京辉与演员黄渤现身杭州西子湖畔,为话剧《活着》举行发布会。据悉,今年9月4日至9月9日,改编自余华同名小说的话剧《活着》将在国家大剧院上演,这也是
近日,由韩寒及独唱团原班人马打造的全新电子读物《一个》在腾讯网上线。腾讯微博邀请原独唱团、现《一个》编辑部全体成员做客,与网友畅谈《一个》之前世今生。韩寒在访谈中
在北京春天的风沙和黄昏的车流里,有时候我会突然想到陆文夫老先生.去世前他常说:“你们要多走出去看一看,再写苏州,写小桥流水,就不是以前那样了。”
近日,中国作家协会在北京顺利举行网络义学作品研讨会,研讨事晓敏的《遍地狼烟》、天下归元的《扶摇皇后》、酒徒的《隋乱》、阿越的《新宋》、杨荟莹的《凝暮颜》5部网络文学