中国人兽共患病学报2011，Vol.27Issue(6)：506-510,5.
A组牛轮状病毒基因工程亚单位疫苗的初步研究
Genetic engineering subunit vaccine of group A bovine rotavirus
摘要
Abstract
In order to acquire rotavirus outer membrane VP4, VP7 and LTB recombinant expression fusion protein and investigate its antigenicity and feasibility as an engineering subunit vaccine of group A bovine rotavirus against calf diarrhea, VP4 and VP7 gene of rotavirus isolate CHLY and LTB from E. coli was cloned and four prokaryotic recombinant expression vectors (pET32a/VP7, pET32a/VP4, pET32a/VP7-LTB and pET32a/VP4-LTB) were constructed and expressed. The recombinant proteins were purified and vaccines were prepared by mixing the protein with Freund's adjuvant. Antigenicity and challenge protection assay of four recombinant proteins were analyzed by immunizing the vaccines on 10w female mice whom were raised with male mice, and the suckling mice whom giving birth to were challenge with 150ul 10-6.5/0.1mL TCID50CHLY isolates. BandScan software analysis result showed that the ratio of rVP7, rVP4, rVP7-LTB and rVP4-LTB to total protein expressed by BL21 (DE3) were 35％, 20％, 30％ and 18％, respectively. Statistical analysis of ten week female mice after immunity revealed that IgG antibody tilter of rVP4-LTB group was highest in the four group. The second one was rVP4 group, the third was rVP7-LTB group, and the forth was rVP7 group. The difference among the four experimental groups were not significant (P＞0.05), but the difference between experimental group and control group was most significant (P＜0.01). Challenging experiment of suckling mice showed that diarrhea ratio of immune group was 13.35％-23.8％, diarrhea duration time was 48h-72h, which was lower than control group. The diarrhea ratio was 100％ and diarrhea duration time was 96h. Challenge protective ratio of four recombinant protein groups were 76.2％-86.7％, among which rVP7-LTB and rVP4-LTB group were a little higher than rVP7 and rVP4 group, but the difference was not significant (P＞0. 05). The experiment illustrated that the four recombinant proteins had better immunogenicity and the neutralization antibody of immune female mice could pass to its offspring and provide protection against rotavirus.关键词
牛轮状病毒/基因工程亚单位疫苗/LTBKey words
bovine rotavirus/ genetic engineering subunit vaccine/ LTB分类
医药卫生引用本文复制引用
杨少华,何洪彬,杨宏军,王长法,高运东,马跃宁,仲跻峰..A组牛轮状病毒基因工程亚单位疫苗的初步研究[J].中国人兽共患病学报,2011,27(6):506-510,5.基金项目
山东省自然科学基金(ZR2009DQ016)资助 （ZR2009DQ016）
