In order to quantitate the bovine immunodeficiency virus line （BIVL） was established by transfecting baby hamster kidney （BIV） cells infection in vitro, a BIV indicator cell with reporter plasmids containing the f...In order to quantitate the bovine immunodeficiency virus line （BIVL） was established by transfecting baby hamster kidney （BIV） cells infection in vitro, a BIV indicator cell with reporter plasmids containing the firefly luciferase gene driven by a BIV long terminal repeat promoter. The BIV activates promoter activity of the LTR to express luciferase upon infection. BIV infection could therefore by quantified by detection of luciferase activity. Compared to standard assays used to detect BIV infection, the BIVL-based assay is 10 times more sensitive than the the CPE-based assay, and has similar sensitivity with the viral capsid protein Western blot assay BIV indicator cell line could detect BIV infection specifically. Luciferase activity of BIV infected BIVL cells showed a time dependent manner, and 60 h post infection is the optimal time to detect BIV infection. Luciferase activity of BIVL cells correlates with the BIV capsid protein expression. Moreover, a linear relationship was found between MOI and the activated intensity of luciferase expression. In brief, the BIV indicator cell line is an easy, robust and quantitive method for monitoring BIV infection.展开更多
The assays for bovine immunodeficiency virus (BIV) induced syncytium formation and BIV long terminal repeat (LTR) directed luciferase (Luc) gene expression were applied to screen and evaluate anti AIDS drugs. Frequen...The assays for bovine immunodeficiency virus (BIV) induced syncytium formation and BIV long terminal repeat (LTR) directed luciferase (Luc) gene expression were applied to screen and evaluate anti AIDS drugs. Frequency of the syncytium formation and BIV LTR directed Luc activity were in proportion to the number of input BIV infected cells. AZT inhibited the syncytium formation and the BIV LTR directed Luc gene expression level. Its inhibitory effects were dosedependent with the IC 50 being 0.24 and 0.052 mmol / L, respectively.展开更多
Transactivator (Tat) of bovine immunodeficiency virus (BIV) is a virus encoded regulatory protein, which activates gene expression directed by BIV long terminal repeat (LTR), and plays an important role in BIV replica...Transactivator (Tat) of bovine immunodeficiency virus (BIV) is a virus encoded regulatory protein, which activates gene expression directed by BIV long terminal repeat (LTR), and plays an important role in BIV replicative cycle. With methods of fusion protein expression and deletion mutation, a set of BIV Tat deletion mutants was constructed, and co transfected into FBL cells with BIV LTR. Using luciferase gene as a reporter, the function of BIV Tat mutants was detected and it showed that: 17/96 aa region of BIV Tat peptide contains full activity; auxiliary amino acids exist in N terminal 17/34 aa region; Cys rich region and basic region are essential to BIV Tat activity. Computer prediction of BIV Tat secondary structure was showed, and mechanism how it functions was analyzed.展开更多
基金The General Foundation of Tianjin Science Committee for Applied Basic Research (08JCZDJC21000)Chinese Ministry of Education (30770081)
文摘In order to quantitate the bovine immunodeficiency virus line （BIVL） was established by transfecting baby hamster kidney （BIV） cells infection in vitro, a BIV indicator cell with reporter plasmids containing the firefly luciferase gene driven by a BIV long terminal repeat promoter. The BIV activates promoter activity of the LTR to express luciferase upon infection. BIV infection could therefore by quantified by detection of luciferase activity. Compared to standard assays used to detect BIV infection, the BIVL-based assay is 10 times more sensitive than the the CPE-based assay, and has similar sensitivity with the viral capsid protein Western blot assay BIV indicator cell line could detect BIV infection specifically. Luciferase activity of BIV infected BIVL cells showed a time dependent manner, and 60 h post infection is the optimal time to detect BIV infection. Luciferase activity of BIVL cells correlates with the BIV capsid protein expression. Moreover, a linear relationship was found between MOI and the activated intensity of luciferase expression. In brief, the BIV indicator cell line is an easy, robust and quantitive method for monitoring BIV infection.
文摘The assays for bovine immunodeficiency virus (BIV) induced syncytium formation and BIV long terminal repeat (LTR) directed luciferase (Luc) gene expression were applied to screen and evaluate anti AIDS drugs. Frequency of the syncytium formation and BIV LTR directed Luc activity were in proportion to the number of input BIV infected cells. AZT inhibited the syncytium formation and the BIV LTR directed Luc gene expression level. Its inhibitory effects were dosedependent with the IC 50 being 0.24 and 0.052 mmol / L, respectively.
文摘Transactivator (Tat) of bovine immunodeficiency virus (BIV) is a virus encoded regulatory protein, which activates gene expression directed by BIV long terminal repeat (LTR), and plays an important role in BIV replicative cycle. With methods of fusion protein expression and deletion mutation, a set of BIV Tat deletion mutants was constructed, and co transfected into FBL cells with BIV LTR. Using luciferase gene as a reporter, the function of BIV Tat mutants was detected and it showed that: 17/96 aa region of BIV Tat peptide contains full activity; auxiliary amino acids exist in N terminal 17/34 aa region; Cys rich region and basic region are essential to BIV Tat activity. Computer prediction of BIV Tat secondary structure was showed, and mechanism how it functions was analyzed.
