摘要
目的克隆并表达发生长片段缺失的HBV核心蛋白（HBV-C）基因，并对其进行DNA序列、蛋白质结构和抗原性分析。方法采用PCR方法从1株野生型HBV基因组中扩增得到发生长片段缺失的HBV-C基因，克隆至pUCm—T质粒，进行测序、同源性比较和蛋白质结构分析；再将基因编码区克隆至原核表达载体pET-28a，构建含HBVC基因的重组表达质粒，转化至大肠埃希菌BL21中进行诱导表达并检测其抗原性。结果PCR扩增出的HBV-C基因长度经序列分析表明，其核苷酸序列缺失了220bp至317bp之间的98个碱基，造成从第74个氨基酸起发生移码突变并失去了抗原性。结论成功克隆和表达了发生长片段缺失的HBVC基因。
Objective and to perform DNA sequen （HBV-C） gene with long fr strain of wild type HBV, w of homology and protein st To clone and express mutant core protein of hepatitis B virus （HBV）, cing, protein structure and antigenicity analyses. Methods The HBV core agment deletion was amplified by polymerase chain reaction （PCR） from a hich was cloned into pUCm-T vector. Then DNA sequencing and analysis ructure were performed. In addition, inserted into the expression constructed, which was then analysis. Results The fragme the sequence, which resulted vector pET-28a and the recombin the coding region of HBV-C was ant plasmid pET-28a-HBV-C was transformed into E. coli BL21 for expression and the antigenicity nt amplified by PCR was 98 hp deletion hetween 220 hp and 317 hp of in frame shift mutation from amino acid 74 and loss of antigenicity. Conclusion The HBV-C gene with long fragment deletion is cloned and expressed successfully.
出处
《中华传染病杂志》
CAS
CSCD
北大核心
2009年第11期650-652,共3页
Chinese Journal of Infectious Diseases
