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Pengaruh Internal Ribosomal Entry Site HIV-1 Terhadap Peningkatan Ekspresi DNA Eksogen Pada Monosit Manusia In Vitro = Effect of HIV-1 Internal Ribosomal Entry Site on Increase of Exogenous DNA Expression in Human Monocytes In Vitro

Main Author: Cintera Rahmagiarti
Format: Book xv, 72 hlm., 21cm x 30cm
Bahasa: ind
Terbitan: Program Magister Ilmu Biomedik , 2018
Subjects:
exogenous DNA
IRES HIV-1
eGFP
Monocytes
NONE
Online Access: http://perpustakaan.fk.ui.ac.id/new-opac/index.php?p=show_detail&id=23887
ctrlnum slims-23887
fullrecord <?xml version="1.0"?> <dc schemaLocation="http://www.openarchives.org/OAI/2.0/oai_dc/ http://www.openarchives.org/OAI/2.0/oai_dc.xsd"><title>Pengaruh Internal Ribosomal Entry Site HIV-1 Terhadap Peningkatan Ekspresi DNA Eksogen Pada Monosit Manusia In Vitro = Effect of HIV-1 Internal Ribosomal Entry Site on Increase of Exogenous DNA Expression in Human Monocytes In Vitro.</title><creator>Cintera Rahmagiarti</creator><subject>exogenous DNA</subject><subject>IRES HIV-1</subject><subject>eGFP</subject><subject>Monocytes</subject><publisher>Program Magister Ilmu Biomedik</publisher><date>2018</date><language>ind</language><type>Book:Book</type><identifier>http://perpustakaan.fk.ui.ac.id/new-opac/index.php?p=show_detail&amp;id=23887</identifier><identifier>T18054fk</identifier><description>DNA eksogen merupakan DNA asing yang diintroduksi ke dalam sel dan sebagai dasar pengembangan vaksin DNA. Ekspresi gen pada DNA eksogen masih rendah dalam antigen-presenting cell (APC) yang merupakan target utama dalam penerapan vaksin DNA, seperti monosit. Salah satu cara dalam meningkatkan ekspresi gen pada DNA eksogen adalah menyisipi sekuen internal inisiasi translasi yaitu IRES HIV-1. Oleh karena itu, pada penelitian ini dilakukan penambahan sekuen IRES HIV-1 pada hulu gen DNA eksogen yang akan diekspresikan di sel monosit manusia. DNA IRES HIV-1_eGFP diamplifikasi dari pcDNA5FRT/TO dan disubkloning ke pcDNA3.1(+). DNA ditransfeksi ke kultur primer monosit dari darah manusia sehat. Sel berfluoresen, persentase sel, dan intensitas fluoresen diamati masing-masing dengan mikroskop fluoresen, Tali cytometer, dan Glomax. Plasmid pcDNA3.1_IRES HIV-1_eGFP berhasil dikonstruksi dan gen egfp berhasil diekspresikan pada sel monosit manusia. Persentase monosit berfluoresen dibandingkan sel kontrol meningkat dari 5% menjadi 11,54% (kelompok pcDNA3.1_eGFP) dan 12,9% (kelompok pcDNA3.1_IRES HIV-1_eGFP) (p=0,297). Intensitas fluoresen eGFP pada kelompok dengan IRES HIV-1 dalam total sel monosit (2,33) dan per monosit (0,069) meningkat signifikan dibandingkan kelompok pcDNA3.1_eGFP dalam total sel monosit (0,08) dan per monosit (0,001) (p=0,001). Oleh karena itu, penambahan IRES HIV-1 terbukti mampu meningkatkan ekspresi gen pada sel monosit secara signifikan. Kata kunci : DNA eksogen, IRES HIV-1, monosit, eGFPExogenous DNA is an transient DNA that is introduced into cells and as a basis for developing DNA vaccines. Gene expression in exogenous DNA is still low in antigen-presenting cells (APC), which is a major target in the application of DNA vaccines, such as monocytes. One way to increase the expression of gene in exogenous DNA is to insert the internal sequence of translation initiation such as IRES HIV-1. Therefore, in this research, the addition of IRES HIV-1 sequence in the upstream gene of exogenous DNA to be expressed in human monocytes cells. IRES HIV-1_eGFP amplified from pcDNA5FRT/TO and subcloned to pcDNA3.1(+).. DNA were transfected into the primary culture of monocytes from healthy human blood. Fluorescent cells, cell percentages, and fluorescent intensity were observed with fluorescent microscope, Tali cytometer, and Glomax respectively. The pcDNA3.1_IRES HIV-1_eGFP successfully constructed and transfected in human monocytes cells. The percentage of fluorescent monocytes compared with control cells increased from 5% to 11.54% (pcDNA3.1_eGFP group) and 12.9% (pcDNA3.1_IRES HIV-1_eGFP) (p = 0.297). The intensity of fluorescent eGFP in the group with IRES HIV-1 in total monocyte (2.33) and each monocyte (0.069) increased significantly compared to pcDNA3.1_eGFP group in total monocyte cell (0.08) and per monocyte (0.001) (p = 0.001). Therefore, the addition of IRES HIV-1 has been shown to increase gene expression in monocyte cells significantly Key words : exogenous DNA , IRES HIV-1, monocytes, eGFP</description><coverage>Jakarta</coverage><type>Other:xv, 72 hlm., 21cm x 30cm</type><subject>NONE</subject><image>http://perpustakaan.fk.ui.ac.id/new-opac/images/default/image.png</image><recordID>slims-23887</recordID></dc>
language ind
format Book:Book
Book
Other:xv, 72 hlm., 21cm x 30cm
Other
author Cintera Rahmagiarti
title Pengaruh Internal Ribosomal Entry Site HIV-1 Terhadap Peningkatan Ekspresi DNA Eksogen Pada Monosit Manusia In Vitro = Effect of HIV-1 Internal Ribosomal Entry Site on Increase of Exogenous DNA Expression in Human Monocytes In Vitro
publisher Program Magister Ilmu Biomedik
publishDate 2018
topic exogenous DNA
IRES HIV-1
eGFP
Monocytes
NONE
url http://perpustakaan.fk.ui.ac.id/new-opac/index.php?p=show_detail&id=23887
contents DNA eksogen merupakan DNA asing yang diintroduksi ke dalam sel dan sebagai dasar pengembangan vaksin DNA. Ekspresi gen pada DNA eksogen masih rendah dalam antigen-presenting cell (APC) yang merupakan target utama dalam penerapan vaksin DNA, seperti monosit. Salah satu cara dalam meningkatkan ekspresi gen pada DNA eksogen adalah menyisipi sekuen internal inisiasi translasi yaitu IRES HIV-1. Oleh karena itu, pada penelitian ini dilakukan penambahan sekuen IRES HIV-1 pada hulu gen DNA eksogen yang akan diekspresikan di sel monosit manusia. DNA IRES HIV-1_eGFP diamplifikasi dari pcDNA5FRT/TO dan disubkloning ke pcDNA3.1(+). DNA ditransfeksi ke kultur primer monosit dari darah manusia sehat. Sel berfluoresen, persentase sel, dan intensitas fluoresen diamati masing-masing dengan mikroskop fluoresen, Tali cytometer, dan Glomax. Plasmid pcDNA3.1_IRES HIV-1_eGFP berhasil dikonstruksi dan gen egfp berhasil diekspresikan pada sel monosit manusia. Persentase monosit berfluoresen dibandingkan sel kontrol meningkat dari 5% menjadi 11,54% (kelompok pcDNA3.1_eGFP) dan 12,9% (kelompok pcDNA3.1_IRES HIV-1_eGFP) (p=0,297). Intensitas fluoresen eGFP pada kelompok dengan IRES HIV-1 dalam total sel monosit (2,33) dan per monosit (0,069) meningkat signifikan dibandingkan kelompok pcDNA3.1_eGFP dalam total sel monosit (0,08) dan per monosit (0,001) (p=0,001). Oleh karena itu, penambahan IRES HIV-1 terbukti mampu meningkatkan ekspresi gen pada sel monosit secara signifikan. Kata kunci : DNA eksogen, IRES HIV-1, monosit, eGFPExogenous DNA is an transient DNA that is introduced into cells and as a basis for developing DNA vaccines. Gene expression in exogenous DNA is still low in antigen-presenting cells (APC), which is a major target in the application of DNA vaccines, such as monocytes. One way to increase the expression of gene in exogenous DNA is to insert the internal sequence of translation initiation such as IRES HIV-1. Therefore, in this research, the addition of IRES HIV-1 sequence in the upstream gene of exogenous DNA to be expressed in human monocytes cells. IRES HIV-1_eGFP amplified from pcDNA5FRT/TO and subcloned to pcDNA3.1(+).. DNA were transfected into the primary culture of monocytes from healthy human blood. Fluorescent cells, cell percentages, and fluorescent intensity were observed with fluorescent microscope, Tali cytometer, and Glomax respectively. The pcDNA3.1_IRES HIV-1_eGFP successfully constructed and transfected in human monocytes cells. The percentage of fluorescent monocytes compared with control cells increased from 5% to 11.54% (pcDNA3.1_eGFP group) and 12.9% (pcDNA3.1_IRES HIV-1_eGFP) (p = 0.297). The intensity of fluorescent eGFP in the group with IRES HIV-1 in total monocyte (2.33) and each monocyte (0.069) increased significantly compared to pcDNA3.1_eGFP group in total monocyte cell (0.08) and per monocyte (0.001) (p = 0.001). Therefore, the addition of IRES HIV-1 has been shown to increase gene expression in monocyte cells significantly Key words : exogenous DNA , IRES HIV-1, monocytes, eGFP
id IOS16788.slims-23887
institution Fakultas Kedokteran Universitas Indonesia
affiliation hellis.onesearch.id
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collection Katalog Perpustakaan Fakultas Kedokteran Universitas Indonesia
repository_id 16788
city JAKARTA PUSAT
province DKI JAKARTA
repoId IOS16788
first_indexed 2022-03-16T03:42:48Z
last_indexed 2022-03-16T03:42:48Z
recordtype dc
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