[The cloning and expression of the truncated human bid gene and its pro-apoptotic effect on Hela cells].
OBJECTIVE
To explore the expression of the truncated bid gene and its pro-apoptotic effect on Hela cells.


METHODS
A full-length human bid gene was cloned by RT-PCR and confirmed by sequence analysis. By deleting the 60 amino acids of N-terminal the truncated bid (tbid) gene was obtained and then inserted into the eukaryotic expression vector pIRES2-EGFP. After the tbid gene was transfected into Hela cells under lipofectamine mediation, the effect of target gene expression on morphology and growth of Hela cells were observed under fluorescence and electron microscopes and analysed by TUNEL staining.


RESULTS
pIRES2-EGFP containing tbid gene was constructed successfully. After Hela cells were transfected with GFP expression vector of tbid gene, tbid was expressed which was followed by decreased cell fluorescence intensity, poor cell growth, and cell death. Cell shrinkage and nuclear condensation, typical apoptotic characteristics, were observed by electron microscope observation and Tunel staining.


CONCLUSIONS
The expression of tbid can effectively induce apoptosis of Hela cells.
Qiu
Xiu-chun
XC
Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China. qiuxiuchun@hotmail.com
Yu
Cui-juan
CJ
Meng
Yan-ling
YL
Xu
Yan-ming
YM
Zhao
Jing
J
Jin
Ming
M
Wang
Cheng-ji
CJ
Fan
Qing-yu
QY
Yang
An-gang
AG
D017209:Apoptosis; D051023:BH3 Interacting Domain Death Agonist Protein; D002352:Carrier Proteins / Q000235:genetics* / Q000502:physiology; D002455:Cell Division; D003001:Cloning, Molecular; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D020287:In Situ Nick-End Labeling; D008164:Luminescent Proteins / Q000235:genetics; D016133:Polymerase Chain Reaction; D014162:Transfection
D051023:BH3 Interacting Domain Death Agonist Protein; C494814:BID protein, human; D002352:Carrier Proteins; D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins