中国畜牧兽医2016，Vol.43Issue(11)：2826-2833,8.DOI:10.16431/j.cnki.1671-7236.2016.11.004
猫重组变应原Fel d 1 与乙肝病毒核心抗原融合基因的原核表达
Prokaryotic Expression of Fusion Gene of Cat Recombinant Allergen Fel d 1 with Hepatitis B Core Antigen
摘要
Abstract
To expose the cat recombinant allergen Fel d 1 protein on the outer surface of hepatitis B core antigen (HBcAg) virus-like particles (VLPs), the recombinant Fel d 1 (rFel d 1) was created by linking the two genes chain 1 and chain 2 that composed the Fel d 1 protein.Then the rFel d 1 sequence was inserted into the HBcAg c/e1 loop area, replacing the amino acids between D78 and E83 in HBcAg c/e1 loop area.We successfully constructed the prokaryotic expression vector pET28a-HBcAg-rFel d 1 via gene codon optimization and synthesis.The recombinant plasmid pET28a-HBcAg-rFel d 1 was transformed into E.coli BL21(DE3) cells, then induced by IPTG, purified by Ni-NTA affinity chromatography and tested by SDS-PAGE, Western blotting and transmission electron microscopy (TEM).The fusion protein HBcAg-rFel d 1 was expressed successfully in E.coli expression system and the pure fusion protein HBcAg-rFel d 1 was purified by Ni-NTA affinity chromatography.Further, TEM confirmed the fusion protein HBcAg-rFel d 1 could assemble into VLPs.The fusion protein HBcAg-rFel d 1 could assemble into VLPs spontaneously, which laid a solid foundation for the research of the preventive and therapeutic vaccines for cat allergy.关键词
猫变应原/乙肝病毒核心抗原/Feld1/病毒样颗粒Key words
cat allergen/hepatitis B virus core antigen/Fel d 1/virus-like particles分类
医药卫生引用本文复制引用
裴业春,安晓荣,侯健,陈永福,闫凤祥,关宏,韦双双,王大勇..猫重组变应原Fel d 1 与乙肝病毒核心抗原融合基因的原核表达[J].中国畜牧兽医,2016,43(11):2826-2833,8.基金项目
国家自然科学基金青年项目(31402257) （31402257）
海南省自然科学基金项目(20153084) （20153084）
