摘要
取pZD 8703质粒DNA(本室构建的高表达乙肝核心抗原基因质粒)用Ava Ⅰ酶切开,Bal 31酶消化,加四种核甘酸dNTP、DNA聚合酶大片段酶补平后,用T_4 DNA连接酶连接,再用EcoRI酶消化,回收EcoRI小片段连接到pJLA 503的EcoRI位点,转化受体菌JM103,获得在P_RP_L启动子控制下,由温度调节CIts 857开启和关闭的高表达乙肝e抗原(HBeAg)的重组体。用抗HBe-β)单克隆酶标抗体配套的ELISA双抗体夹心法测定经超声波处理的细菌原液,HBeAg的终效价在1:10240以上,只含少量HBeAg。可以代替血清HBeAg用作配套检测试剂。
pZD 8703 DNA was digested with restriction endonuclease AvaI and nuclease Bal 31, addedd NTP, polymerase Ⅰ (L. F.), recombinant with pJLA 503 EcoRI fragment, transformated into E. coli JM 103. E. coli pZD 8901, a strain of high expressing HBeAg was obtained. A series of testing results verified that the produced protein of E. coli pZD 8901 has a good charactristics for clinical ELISA detecting kits of hepatitis B.
出处
《中国药科大学学报》
CAS
CSCD
北大核心
1992年第5期290-293,共4页
Journal of China Pharmaceutical University
