Cloning and eukaryotic expression of HIV-1B gp120 genes from a patient with AIDS dementia complex
10.3760/cma.j.issn.1003-9279.2012.02.005
- VernacularTitle:艾滋病痴呆综合征患者HIV-1B gp120基因的克隆及真核表达
- 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
1. 山东大学
- Keywords:
Acquired immunodeficiency syndrome;
HIV-1;
Cloning,molecular;
Expressedsequence tags
- From:
Chinese Journal of Experimental and Clinical Virology
2012;26(2):93-95
- CountryChina
- Language:Chinese
-
Abstract:
Objective To clone and express the HIV-1B gp120 genes isolated at different organizations from a patient died of AIDS dementia complex (ADC) in eukaryotic cells.Methods Using 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,HIVIB 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.Results HIV-I B 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.i ( + ) could express envelope glycoprotein HIV-1B gp120 in U87 cells.Conclusion All 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.
