AI Article Synopsis
- The GST fusion protein expression system, along with the protein truncation test (PTT) protocol, was employed to identify gene frame shift mutations in the Lis1 gene from hepatocarcinoma samples.
- Cloning the RT-PCR products into a GST expression vector and expressing them in E. coli led to the unexpected production of a truncated 33 kD protein instead of the expected 71 kD full-length protein.
- Sequencing revealed an insertion mutation that caused premature termination of translation, demonstrating that the enhanced PTT assay is a quick and effective method for detecting frame shift mutations in genes.
Article 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.
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