摘要
X基因是乙型肝炎病毒(HBV)的四个开读框架之一,编码一个含154个氨基酸的蛋白。将含有编码145个氨基酸的X基因主要部分用Bam HI及Bgl Ⅱ酶切,获得Bam HI-Bgl Ⅱ HBV DNA片段(584bp),将其插入到质粒pEx31的Bam HI切点,构建成一个在λPL启动子控制下表达MS2-X融合蛋白的质粒—pEx31-HBX。含此质粒的大肠杆菌可产生占菌体蛋白总量40％的28千道尔顿融合蛋白。用抗X标准抗血清证实,此融合蛋白具有X的抗原活性,并制备了抗融合蛋白的抗血清。这是在国内首次成功地高效表达乙型肝炎病毒的X蛋白。
Region X is one of the four open reading frames ( ORFs ) of hepatitis B virus and encodes a polypeptide of 154 amino acids. A 584 bp BamHI-Rgl Ⅱ fragment of the HBV DNA containing the major part of ORF X which encodes the C terminal 145 aminc acids was inserted into the Bam HI site of plasmid pEx31. This insertion resulted in an MS2-polymerase-X fused protein of 243 amino acids under the control of PL promoter. Cells harboring plasmid pEx31-HBX derived from the above construct overproduced a 28 kDa fused protein in E.coli at a level of 40% of total cellular protein. The fused protein was recognized by the anti-X antibodies and had been used as immunogen to produse antisera. This is the first demonstration of high-level production of X protein in E.coli in our country.
出处
《病毒学报》
CAS
CSCD
北大核心
1989年第4期318-322,共5页
Chinese Journal of Virology
基金
国家自然科学基金
