Cytosine deaminase gene therapy for gastrointestinal cancer research.docVIP

 PAGE \* MERGEFORMAT 23 Cytosine deaminase gene therapy for gastrointestinal cancer research Abstract: The cytosine deaminase gene into tumor cells may be non-toxic to the original drug metabolism in tumor cells as the product of a cytotoxic anti-tumor effect to play. In this paper, the gene cloning, mechanism and treatment of gastrointestinal cancer research. With the rapid development of molecular biology, gene therapy methods, although more than this new experimental research tools, they are showing good prospects [1]. Since 1991, Huber et al [2] proposed the original virus directed enzyme prodrug therapy (virus directed enzyme prodrug therapy, VDEPT) has been related to genetic research has become a hot topic. At present, a new gene therapy system --EC-CD/5-FC (Escherichia coli cytosine deaminase / 5 - fluorocytosine) systems are increasingly subject to great attention [3]. In this paper, cytosine deaminase (cytosine deaminase, CD) the status of gene therapy for gastrointestinal tumors are reviewed. Cloning CD E. coli cytosine deaminase is an enzyme in the metabolism of bypass, mammalian cells do not contain the enzyme. Austin et al [4] in 1992, starting with E. coli strain used in the traditional method of extracting plasmid DNA, and further separation, purification, and transfection of host bacteria, initially built into the prokaryotic cells of CD vector, cloning, encoding CD’s E. coli genes, the use of positive genetic screening methods to obtain a carrying 10.8 kb of DNA inserted into plasmid, in which a 3 kb of DNA fragment retains CD activity. CK protein molecular weight of 52000, DNA sequence analysis showed that one contained 427 amino acids open reading frame encoding CD. To confirm that CD gene expression in eukaryotic cells in order to play a role, the researchers used oligonucleotide mutation (oligonucleotide site-directed mutagenesis) technology, the CD gene start codon from GTG → ATG, res