TY  - JOUR
AU  - Liang, C.
AU  - Xu, X.
AU  - Yu, H.
AU  - Zhou, X.
AU  - Wen, X.
AU  - Li, Y.
AU  - Kojić, Snežana
AU  - Wang, Y.
AU  - Ma, G.
PY  - 2019
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1229
AB  - Objective: To construct the recombinant lentiviral vector containing cardiac adriamycin responsive protein (CARP) gene and small-hairpin RNA (shRNA) targeting CARP gene and to pack the lentivirus, and to lay the foundation for further study on the function and mechanism of CARP in adriamycin (ADR) induced cardiomyopathy. Methods: After the rat CARP gene was amplified by PCR and shRNAs targeting CARP were designed and synthesized, they were inserted into the shuttle plasmids GV-358 or GV-248. respectively. After confirmed by sequencing, the recombinant shuttle vectors containing CARP gene or shRNA and auxiliary packaging plasmid Helper 1 0 and Helper 2 0 were co transfected into the IIEK293T cells for virus packaging and amplification; the viral titer was detected by end point dilution. The IIEK293T cells were infected with the recombinant lentiviruses. and the green fluoresence intensity was observed by fluorescence mircroscope. The H9C2 cells were infected with the recombinant lentivirues and divided into control group. CARP overexpression group and shRNA group; Western blotting method was used to detect the CARP expression levels in the cells in various groups. Results: The DNA sequencing results showed that the sequences of CARP overexpression and shRNA vectors were in accordance with the designed sequences. The expression of green fluorescence protein was seen under fluorescence microscope after transfection of the vectors of the IIEK293T cells. After infection of the H9C2 cells, the expression level of CARP protein in CARP overexpression group was 5. 3 times higher than that in control group ( P=0. 01); while it was down-regulated by 53% in CARP shRNA group compared with control group ( P= 0 02). Conclusion: The lentivirus expression vectors carrying CARP or shRNA targeting CARP are successfully constructed and the lentiviruses obtained could significantly interfere the expression of CARP in the II9C2 cells.
PB  - Jilin University Press
T2  - Journal of Jilin University Medicine Edition
T1  - Construction and identification of recombinant lentivirus overexpression and RNA interference vector containing cardiac adriamycin reactive protein gene
EP  - 771
IS  - 4
SP  - 766
VL  - 45
DO  - 10.13481/j.1671-587x.20190404
ER  - 

@article{
author = "Liang, C. and Xu, X. and Yu, H. and Zhou, X. and Wen, X. and Li, Y. and Kojić, Snežana and Wang, Y. and Ma, G.",
year = "2019",
abstract = "Objective: To construct the recombinant lentiviral vector containing cardiac adriamycin responsive protein (CARP) gene and small-hairpin RNA (shRNA) targeting CARP gene and to pack the lentivirus, and to lay the foundation for further study on the function and mechanism of CARP in adriamycin (ADR) induced cardiomyopathy. Methods: After the rat CARP gene was amplified by PCR and shRNAs targeting CARP were designed and synthesized, they were inserted into the shuttle plasmids GV-358 or GV-248. respectively. After confirmed by sequencing, the recombinant shuttle vectors containing CARP gene or shRNA and auxiliary packaging plasmid Helper 1 0 and Helper 2 0 were co transfected into the IIEK293T cells for virus packaging and amplification; the viral titer was detected by end point dilution. The IIEK293T cells were infected with the recombinant lentiviruses. and the green fluoresence intensity was observed by fluorescence mircroscope. The H9C2 cells were infected with the recombinant lentivirues and divided into control group. CARP overexpression group and shRNA group; Western blotting method was used to detect the CARP expression levels in the cells in various groups. Results: The DNA sequencing results showed that the sequences of CARP overexpression and shRNA vectors were in accordance with the designed sequences. The expression of green fluorescence protein was seen under fluorescence microscope after transfection of the vectors of the IIEK293T cells. After infection of the H9C2 cells, the expression level of CARP protein in CARP overexpression group was 5. 3 times higher than that in control group ( P=0. 01); while it was down-regulated by 53% in CARP shRNA group compared with control group ( P= 0 02). Conclusion: The lentivirus expression vectors carrying CARP or shRNA targeting CARP are successfully constructed and the lentiviruses obtained could significantly interfere the expression of CARP in the II9C2 cells.",
publisher = "Jilin University Press",
journal = "Journal of Jilin University Medicine Edition",
title = "Construction and identification of recombinant lentivirus overexpression and RNA interference vector containing cardiac adriamycin reactive protein gene",
pages = "771-766",
number = "4",
volume = "45",
doi = "10.13481/j.1671-587x.20190404"
}

Liang, C., Xu, X., Yu, H., Zhou, X., Wen, X., Li, Y., Kojić, S., Wang, Y.,& Ma, G.. (2019). Construction and identification of recombinant lentivirus overexpression and RNA interference vector containing cardiac adriamycin reactive protein gene. in Journal of Jilin University Medicine Edition
Jilin University Press., 45(4), 766-771.
https://doi.org/10.13481/j.1671-587x.20190404

Liang C, Xu X, Yu H, Zhou X, Wen X, Li Y, Kojić S, Wang Y, Ma G. Construction and identification of recombinant lentivirus overexpression and RNA interference vector containing cardiac adriamycin reactive protein gene. in Journal of Jilin University Medicine Edition. 2019;45(4):766-771.
doi:10.13481/j.1671-587x.20190404 .

Liang, C., Xu, X., Yu, H., Zhou, X., Wen, X., Li, Y., Kojić, Snežana, Wang, Y., Ma, G., "Construction and identification of recombinant lentivirus overexpression and RNA interference vector containing cardiac adriamycin reactive protein gene" in Journal of Jilin University Medicine Edition, 45, no. 4 (2019):766-771,
https://doi.org/10.13481/j.1671-587x.20190404 . .

TY  - JOUR
AU  - Jha, S.S.
AU  - Danelishvili, L.
AU  - Wagner, D.
AU  - Maser, J.
AU  - Li, Y.-J.
AU  - Morić, Ivana
AU  - Vogt, S.
AU  - Yamazaki, Y.
AU  - Lai, B.
AU  - Bermudez, L.E.
PY  - 2010
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/469
AB  - Background: Mycobacterium avium subsp hominissuis (previously Mycobacterium avium subsp avium) is an environmental organism associated with opportunistic infections in humans. Mycobacterium hominissuis infects and replicates within mononuclear phagocytes. Previous study characterized an attenuated mutant in which the PPE gene (MAV-2928) homologous to Rv1787 was inactivated. This mutant, in contrast to the wild-type bacterium, was shown both to have impaired the ability to replicate within macrophages and to have prevented phagosome/lysosome fusion. Results: MAV-2928 gene is primarily upregulated upon phagocytosis. The transcriptional profile of macrophages infected with the wild-type bacterium and the mutant were examined using DNA microarray, which showed that the two bacteria interact uniquely with mononuclear phagocytes. Based on the results, it was hypothesized that the phagosome environment and vacuole membrane of the wild-type bacterium might differ from the mutant. Wild-type bacterium phagosomes expressed a number of proteins different from those infected with the mutant. Proteins on the phagosomes were confirmed by fluorescence microscopy and Western blot. The environment in the phagosome of macrophages infected with the mutant differed from the environment of vacuoles with M. hominissuis wild-type in the concentration of zinc, manganese, calcium and potassium. Conclusion: The results suggest that the MAV-2928 gene/operon might participate in the establishment of bacterial intracellular environment in macrophages.
PB  - BMC
T2  - BMC Microbiology
T1  - Virulence-related Mycobacterium avium subsp hominissuis MAV-2928 gene is associated with vacuole remodeling in macrophages
VL  - 10
DO  - 10.1186/1471-2180-10-100
ER  - 

@article{
author = "Jha, S.S. and Danelishvili, L. and Wagner, D. and Maser, J. and Li, Y.-J. and Morić, Ivana and Vogt, S. and Yamazaki, Y. and Lai, B. and Bermudez, L.E.",
year = "2010",
abstract = "Background: Mycobacterium avium subsp hominissuis (previously Mycobacterium avium subsp avium) is an environmental organism associated with opportunistic infections in humans. Mycobacterium hominissuis infects and replicates within mononuclear phagocytes. Previous study characterized an attenuated mutant in which the PPE gene (MAV-2928) homologous to Rv1787 was inactivated. This mutant, in contrast to the wild-type bacterium, was shown both to have impaired the ability to replicate within macrophages and to have prevented phagosome/lysosome fusion. Results: MAV-2928 gene is primarily upregulated upon phagocytosis. The transcriptional profile of macrophages infected with the wild-type bacterium and the mutant were examined using DNA microarray, which showed that the two bacteria interact uniquely with mononuclear phagocytes. Based on the results, it was hypothesized that the phagosome environment and vacuole membrane of the wild-type bacterium might differ from the mutant. Wild-type bacterium phagosomes expressed a number of proteins different from those infected with the mutant. Proteins on the phagosomes were confirmed by fluorescence microscopy and Western blot. The environment in the phagosome of macrophages infected with the mutant differed from the environment of vacuoles with M. hominissuis wild-type in the concentration of zinc, manganese, calcium and potassium. Conclusion: The results suggest that the MAV-2928 gene/operon might participate in the establishment of bacterial intracellular environment in macrophages.",
publisher = "BMC",
journal = "BMC Microbiology",
title = "Virulence-related Mycobacterium avium subsp hominissuis MAV-2928 gene is associated with vacuole remodeling in macrophages",
volume = "10",
doi = "10.1186/1471-2180-10-100"
}

Jha, S.S., Danelishvili, L., Wagner, D., Maser, J., Li, Y.-J., Morić, I., Vogt, S., Yamazaki, Y., Lai, B.,& Bermudez, L.E.. (2010). Virulence-related Mycobacterium avium subsp hominissuis MAV-2928 gene is associated with vacuole remodeling in macrophages. in BMC Microbiology
BMC., 10.
https://doi.org/10.1186/1471-2180-10-100

Jha S, Danelishvili L, Wagner D, Maser J, Li Y, Morić I, Vogt S, Yamazaki Y, Lai B, Bermudez L. Virulence-related Mycobacterium avium subsp hominissuis MAV-2928 gene is associated with vacuole remodeling in macrophages. in BMC Microbiology. 2010;10.
doi:10.1186/1471-2180-10-100 .

Jha, S.S., Danelishvili, L., Wagner, D., Maser, J., Li, Y.-J., Morić, Ivana, Vogt, S., Yamazaki, Y., Lai, B., Bermudez, L.E., "Virulence-related Mycobacterium avium subsp hominissuis MAV-2928 gene is associated with vacuole remodeling in macrophages" in BMC Microbiology, 10 (2010),
https://doi.org/10.1186/1471-2180-10-100 . .