Detection of Lis1 Gene Frame
Shift Mutation in Human Hepatocarcinoma
XIA Shuang-Luo, ZHAO Mu-Jun1*, WU Zhen-Yu1,
LI Zai-Ping1
( Department of Biochemical Engineering, School of Life Science, Shanghai
University, Shanghai 201800, China； 1Shanghai Institute of Biochemistry, the
Chinese Academy of Sciences, Shanghai 200031, China )
Abstract GST fusion
protein expression system combined with protein truncation test(PTT) protocol
was used to detect gene frame shift mutation. The RT-PCR products of Lis1
genes from hepatocarcinoma samples were respectively cloned into a GST fusion
protein expression vector pGEX-1, then expressed in E.coli. The
results showed a truncated 33 kD fusion protein in SDS-PAGE, although the
full-translated product of Lis1 gene should be of 71 kD. Sequencing
revealed insertion of an A residue, causing the premature termination of
translation, between the 163th and 164th nucleotide of Lis1 gene. This
improved PTT assay was proved to be a fast and effective way in detecting gene
frame shift mutation.
Key words PTT； Lis1 gene； frame shift mutation； GST fusion protein
*Corresponding author： Tel, 86-21-64374430-295； Fax, 86-21-64338357； e-mail, [email protected]