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利用广宿主范围载体在甲醇醋杆菌中表达乙型肝炎病毒(HBV)核心抗原基因。

Expression of the core antigen gene of hepatitis B virus (HBV) in Acetobacter methanolicus using broad-host-range vectors.

作者信息

Schröder R, Maassen A, Lippoldt A, Börner T, von Baehr R, Dobrowolski P

机构信息

Institut für Biotechnologie, Leipzig, Federal Republic of Germany.

出版信息

Appl Microbiol Biotechnol. 1991 Aug;35(5):631-7. doi: 10.1007/BF00169628.

Abstract

Using the broad-host-range promoter probe vector pRS201 for cloning of phage Acm1 promoters, we established a convenient vector system for expression of heterologous genes in different Gram-negative bacteria. The usefulness of this system was demonstrated by expression of the HBV core gene in Acetobacter methanolicus. Plasmids carrying the HBV core gene downstream of different Acm1-phage promoters were transferred to A. methanolicus, a new potential host for recombinant DNA expression. Using enzyme immunoassay and immunoblot techniques, the amount and composition of core antigen produced in A. methanolicus were compared with that derived from Escherichia coli. The expression of immunoreactive core antigen in A. methanolicus exceeds by sevenfold that in E. coli using an expression system with tandemly arranged promoters. Morphological observations by electron microscopy show that the HBV core gene products isolated from both hosts are assembled into regular spherical particles with a diameter of about 28 nm that are comparable to original viral nucleocapsids.

摘要

利用广宿主范围启动子探针载体pRS201克隆噬菌体Acm1启动子,我们建立了一个便于在不同革兰氏阴性细菌中表达异源基因的载体系统。通过在甲醇醋杆菌中表达乙肝病毒核心基因,证明了该系统的实用性。携带不同Acm1噬菌体启动子下游乙肝病毒核心基因的质粒被转移到甲醇醋杆菌中,甲醇醋杆菌是重组DNA表达的一种新的潜在宿主。利用酶免疫测定和免疫印迹技术,将甲醇醋杆菌中产生的核心抗原的量和组成与大肠杆菌来源的进行了比较。在使用串联排列启动子的表达系统中,甲醇醋杆菌中免疫反应性核心抗原的表达比大肠杆菌中的高出七倍。电子显微镜形态学观察表明,从两种宿主中分离出的乙肝病毒核心基因产物组装成直径约28nm的规则球形颗粒,与原始病毒核衣壳相当。

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