title: Detection of Lis1 Gene Frame Shift Mutation in Human Hepatocarcinoma.
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.
authors.affiliation: Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China. mjzhao@sunm.shcnc.ac.cn
authors.forename: Shuang-Luo
authors.initials: SL
authors.lastname: Xia
authors.forename: Mu-Jun
authors.initials: MJ
authors.lastname: Zhao
authors.forename: Zhen-Yu
authors.initials: ZY
authors.lastname: Wu
authors.forename: Zai-Ping
authors.initials: ZP
authors.lastname: Li