Cloning and eukaryotic expression of HIV-1B gp120 genes from a patient with AIDS dementia complex.
- Author:
Li ZHAO
1
;
Yu-Fen YAN
;
Jing LI
;
Shuang-Shuang PU
;
Zhi-Yu WANG
;
Hong-Ling WEN
;
Yan-Yan SONG
;
Hong-Zhi XU
Author Information
- Publication Type:Journal Article
- MeSH: AIDS Dementia Complex; virology; Cloning, Molecular; HIV Envelope Protein gp120; biosynthesis; genetics; toxicity; Humans; Recombinant Proteins; biosynthesis; Sequence Analysis, DNA
- From: Chinese Journal of Experimental and Clinical Virology 2012;26(2):93-95
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo clone and express the HIV-1B gp120 genes isolated at different organizations from a patient died of AIDS dementia complex (ADC) in eukaryotic cells.
METHODSUsing the genomic DNA isolated from peripheral lymphnodes, choroid plexus and occipital white matter from a patient died of ADC as the template, HIV-1B gp120 gene was amplified with PCR. After sequenced, HIV-1B gp120 was inserted into pcDNA3.1 (+) and recombinant expressing vector gp120/pcDNA3.1 (+) was constructed succeffuly confirming with sequencing. Then expressing vector was transfected into eukaryotic cells U87 using liposome transfection and expression of HIV-1B gp120 gene was assayed with indirect immunofluorescence.
RESULTSHIV-1B gp120 genes isolated from peripheral lymphnodes, choroid plexus and occipital white matter of the ADC patient were successfully cloned and recombinant expressing vector gp120/pcDNA3; 1 (+) could express envelope glycoprotein HIV-1B gp120 in U87 cells.
CONCLUSIONAll the HIV-1B gp120 gene isolated at the different organizations of the same ADC patient could express in U87 cells, which may supply a valuable basis for studying the neurotoxicity and neurotoxic mechanism of HIV-1 gp120 protein.