Establishment of Cell
Line with the FEN-1 Gene Blocked by Its Antisense
SHI Bin-Shan, YU Ying-Nian*, CAI Zhu-Nan
( Department of Pathophysiology, College of Medicine, Zhejiang University,
Hangzhou 310031, China )
SHEN Bin-Hui
( City of Hope National Medical Center and Beckman Research Institute,
Duart, CA91010-3000 USA ）
Abstract FEN-1 is a
structure-specific endo/exonuclease, which is involved in the process of both
DNA duplication and DNA repair. In this work a mammalian expression vector
expressing antisense FEN-1 gene fragment pMAMneoAmp–FNB–
was constructed, after cloning the NcoI-BamHI fragment of FEN-1
gene into the mammalian expression vector pMAMneoAmp– in antisense
orientation. After FL cell was transfected with pMAMneoAmp–FNB–
and selected by G418, the FL-FEN-1– cell line, in which the FEN-1
gene expression was blocked, was established. It was found that the growth of
FL-FEN-1– was decreased upon the induction with dexamethasone and
its TD was 3.03 d, while the TD of controls FL and FL-M
induced with dexamethasone was 2.03 and 2.22 d, respectively, and the TD of the FL-FEN-1–
cell without dexamethasone was 2.38 d.
Key words structure-specific nuclease FEN-1； antisense nucleic acid
technique；
plasmid construction； FL cell
Received: July 23,
1999 Accepted: September 10, 1999
This work was supported by a grant from the National Natural Science Foundation
of China, No. 39830210 and No. 39870340
* Corresponding author: Tel/Fax: 86-571-7217149; e-mail, [email protected]