title: Identification and functional analysis of the GTPV bidirectional promoter region.
abstract: The goat pox chick embryo-attenuated virus (GTPV) has been developed as an effective vaccine that can elicit protective immune responses. It possesses a large genome and a robust ability to express exogenous genes. Thus, this virus is an ideal vector for recombinant live vaccines for infectious diseases in ruminant animals. In this study, we identified a novel bidirectional promoter region of GTPV through screening named PbVV(±). PbVV(±) is located between ETF-l and VITF-3, which are transcribed in opposite directions. A new recombinant goat pox virus (rGTPV) was constructed, in which duplicate PbVV(+) was used as a promoter element to enhance Brucella OMP31 expression, and duplicate PbVV(-) was used as a promoter element to regulate enhanced green fluorescent protein (EGFP) at the same time as the selection marker. PbVV(-) promoter activity was compared to that of the P7.5 promoter of vaccinia virus, as measured by EGFP expression; the fluorescence intensity of EGFP expressed in cells was confirmed by fluorescence microscopy and flow cytometry. PbVV(+) promoter activity was measured by Brucella OMP31 expression. Interaction with the anti-Brucella-OMP31 monoclonal antibody was confirmed by western blotting, and OMP31 mRNA expression was assessed by qRT-PCR. The results of this study will be useful for the further study of effective multivalent vaccines based on rGTPV. This study also provides a theoretical basis for overcoming the problem of low expression of exogenous genes.
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.
authors: Hui
authors: H
authors: Zhang
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.
authors: Zhihua
authors: Z
authors: Sun
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.
authors: Na
authors: N
authors: Zhang
authors: School of Life Sciences, Shangqiu Normal University, Shangqiu, Henan Province, 476000, People's Republic of China.
authors: Zhiqiang
authors: Z
authors: Li
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.
authors: Pengyan
authors: P
authors: Wang
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.
authors: Qiang
authors: Q
authors: Fu
authors: School of Medicine, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.
authors: Yan
authors: Y
authors: Ren
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.
authors: Xuehua
authors: X
authors: Shao
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.
authors: Yu
authors: Y
authors: Zhang
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China.
authors: Zhiru
authors: Z
authors: Guo
authors: College of Animal Science and Technology, Shihezi University, Shihezi, Xinjiang Province, 832000, People's Republic of China. ccf_xb@126.com.
authors: Chuangfu
authors: C
authors: Chen
mesh_terms: D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D018151:Capripoxvirus / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D011401:Promoter Regions, Genetic
chemical_list: D001425:Bacterial Outer Membrane Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: Bidirectional promoter; Goat pox virus; Omp31; Vector