草业学报Issue(4)：138-145,8.DOI:10.11686/cyxb20140417
禽流感血凝素HA超表达载体的构建及高表达百脉根的获得
Vector construction and over-expression of an avian influenza virus hemagglutinin (HA)in transgenic Lotus corniculatus
摘要
Abstract
A safe and efficient plant expression vector,HA-LTB,which added double 35S promoter,Ωenhan-cer,endoplasmic reticulum retention signal KDEL sequence and extended poly (A)sequence was constructed. Twenty one transgenic Lotus corniculatus plants were obtained by the Agrobacterium-mediated method and the results of PCR and RT-PCR showed that the HA-LTB gene had been obtained from the plant genome. ELISA results of the B subunit of Escherichiacoli heat-labile enterotoxin (LTB)showed that LTB could be ex-pressed in transgenic L. corniculatus and had immunogenicity.It was concluded that hemagglutinin (HA ) which was fused with LTB had immune activity.The highest expressions of LTB and HA were 0.086μg/mg (crude protein extracts)and 0.25μg/mg (crude protein extracts)respectively which has approached advanced international standards.A high level exogenous antigen protein expression platform which was a basis for re-search on the H5N1 avian influenza vaccine was established by optimizing the expression vector.关键词
百脉根/载体构建/H A抗原蛋白/LTB免疫佐剂/高表达Key words
Lotus corniculatus/vector construction/hemagglutinin (HA)/LTB/a high expression level分类
农业科技引用本文复制引用
张伟伟,袁蓓,张占路,赵杨敏,徐秉良,吴燕民..禽流感血凝素HA超表达载体的构建及高表达百脉根的获得[J].草业学报,2014,(4):138-145,8.基金项目
国家重点基础研究发展计划“973”项目（2014CB138701）和国家自然科学基金资助项目（31372361）资助。 （）
