[Effect of the truncated human apoptosis-inducing factor expression on apoptosis of HeLa cells].
OBJECTIVE
To observe the expression of the truncated human apoptosis-inducing factor (AIF) gene and its apoptosis-inducing effect on HeLa cells.


METHODS
Full-length human AIF gene was cloned by RT-PCR, then the truncated AIF gene was constructed by deleting the N-terminal mitochondrial location sequence (MLS), and inserted into the EGFP co-expression vector pIRES2-EGFP. After being transfected into HeLa cells with Lipofectamin, the expression of the truncated AIF gene and its effect on HeLa cells morphology and growth condition were detected by fluorescence microscope, immunohistochemical staining, indirect fluoroimmunoassay and electron microscope analysis.


RESULTS
The eukaryotic expression vector pIRES2-EGFP containing of truncated human AIF gene was constructed successfully. The AIF protein could be detected in the transfected HeLa cells. After transfection, typical apoptotic feature of the transfected HeLa cells and a mass of cell death were observed under electron microscope.


CONCLUSIONS
The expression of the truncated human AIF gene can induce apoptosis of the transfected human HeLa cells.
Yu
Cui-juan
CJ
Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China.
Meng
Yan-jing
YJ
Zhao
Jing
J
Wang
Zhi
Z
Wang
Cheng-ji
CJ
Yang
An-gang
AG
D017209:Apoptosis; D051033:Apoptosis Inducing Factor / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D003001:Cloning, Molecular; D005784:Gene Amplification; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D019169:Jurkat Cells; D010446:Peptide Fragments / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D051033:Apoptosis Inducing Factor; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins