Prokaryotic expression, localization and function of the infectious laryngotracheitis virus glycoprotein G  

作　　者：SUN Zhaogang ZHANG Manfu 

机构地区：[1]Lab of Animal Molecular Virology, College of Biological Science, China Agricultural University, Beijing 100094, China [2]Department of Bacteriology and Immunology, Beijing TB & thoracic Tumor Research Institute, Beijing 101149, China

出　　处：《Chinese Science Bulletin》2005年第18期2033-2038,共6页

摘　　要：The infectious laryngotracheitis virus (ILTV) glycoprotein G (gG) gene of E3 and Zhonghai strains was cloned, sequenced and compared with the gG gene of other Type I animal herpesviruses. To find the localization and the function of the gG in the infected cells, the 35 kD fusion pro-tein (His-GG) was expressed by inserting the coding region of gG except for the signal peptide into pET30a (+). After puri-fication of the His-GG fusion protein, the rats’ antibody to the His-GG was prepared and purified by using the protein G Sepharose. Results of laser scanning confocal microscopy (LSCM) detection showed that the ILTV gG was in the peri-nuclear region and membrane of chicken embryo liver (CEL) and kidney (CEK) cells, and that the gG accumulated more in the coalescent part than in the other parts of the adjacent CEL or CEK cells. The plaque size and the one-step growth curve tests suggested that the ILTV gG was required for viral growth by cell-to-cell direct infection in tissue-cultured CEL cells.The infectious laryngotracheitis virus （ILTV） glycoprotein G （gG） gene of E3 and Zhonghai strains was cloned, sequenced and compared with the gG gene of other Type Ⅰ animal herpesviruses. To find the localization and the function of the gG in the infected cells, the 35 kD fusion protein （His-GG） was expressed by inserting the coding region of gG except for the signal peptide into pET30a （＋）. After purification of the His-GG fusion protein, the rats＇ antibody to the His-GG was prepared and purified by using the protein G Sepbarose. Results of laser scanning confocal microscopy （LSCM） detection showed that the ILTV gG was in the perinuclear region and membrane of chicken embryo liver （CEL） and kidney （CEK） cells, and that the gG accumulated more in the coalescent part than in the other parts of the adjacent CEL or CEK cells. The plaque size and the one-step growth curve tests suggested that the ILTV gG was required for viral growth by cell-to-cell direct infection in tissue-cultured CEL cells.

关 键 词：原核表达 糖蛋白 传染性 喉气管炎 缩氨酸 

分 类 号：S852.65[农业科学—基础兽医学]