title: A simple selection system for construction of recombinant gD-negative pseudorabies virus as a vaccine vector.
abstract: We describe a simple, efficient two-step method for construction of glycoprotein D (gD)-negative pseudorabies virus (PrV) carrying transgenes inserted in place of the gD gene. The first step was the use of the thymidine kinase (TK) gene of herpes simplex virus (HSV) for insertional inactivation of the gD gene in a PrV mutant deficient in both TK and glycoprotein E (gE). The gD-negative, HSV-TK-positive mutant could be selected in HAT medium. The second step was substitution of HSV-TK with other genes of interest. The resultant gD/gE/TK-negative mutant was easily isolated by acyclovir selection. The expression of the transgene was detectable in vivo and the antibody responses against both inserted antigens and PrV were induced. The protective efficacy of the gD/gE/TK-negative PrV against lethal PrV challenge was also demonstrated. This PrV mutant carrying immunogenic proteins from unrelated porcine pathogens may be tested as a multivalent vaccine candidate for swine.
authors.affiliation: Department of Microbiology, National Cheng Kung University Medical College, 1 Dashiue Road, 701, Tainan, Taiwan.
authors.forename: Ai Li
authors.initials: AL
authors.lastname: Shiau
authors.forename: Chia Wen
authors.initials: CW
authors.lastname: Liu
authors.forename: Shiang Yiu
authors.initials: SY
authors.lastname: Wang
authors.forename: Chiau Yuang
authors.initials: CY
authors.lastname: Tsai
authors.forename: Chao Liang
authors.initials: CL
authors.lastname: Wu
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis; D002460:Cell Line; D005822:Genetic Vectors; D011558:Herpesvirus 1, Suid / Q000235:genetics* / Q000276:immunology; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D018482:Muscle, Skeletal / Q000378:metabolism; D018139:Simplexvirus / Q000201:enzymology; D013552:Swine; D013937:Thymidine Kinase / Q000235:genetics*; D019076:Transgenes; D014614:Vaccines, Synthetic / Q000096:biosynthesis*; D014759:Viral Envelope Proteins / Q000235:genetics*
chemical_list: D000914:Antibodies, Viral; D014614:Vaccines, Synthetic; D014759:Viral Envelope Proteins; C042966:pseudorabies virus glycoproteins; D013937:Thymidine Kinase