Engineer Novel Anticancer Bioagents
Abstract
During the second contract-year of our research we have successfully reconstituted the entire FK228 biosynthetic gene cluster (~35 kb) plus additional DNA elements (total ~6 kb) into a single integrative vector, a miniHimar RB1 transposon, through complicated manipulations. However, repeated attempts to integrate the gene cluster-carrying transposon into E. coli chromosome for stable functioning without antibiotic selection (hence to create marker-free genetically modified organism--GMO as required by FDA regulations) have failed. The overall transformation efficiency is extremely low (<10-9 transformants per ug DNA) and always resulted in partial deletion or rearrangement of the gene cluster. We reason that the large size of the gene cluster (total 38 kb, which is still one of the small gene clusters known to produce an FDA-approved anticancer drug) and two internal homologous regions might have contributed to the problem, which was not forseen in the first place. To overcome those problems, we have designed and initiated a completely new strategy which employs a much small gene cluster (~9 kb) that contains only two genes, for the production of a potent anticancer agent tryprostatin B. We are therefore behind the projected schedule of work and we modified the project design. We are confident that we can achieve the project goal but we one year of non-budgetary extension of the entire project duration to end by 09/14/2012.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2010
- Accession Number
- ADA535365
Entities
People
- Cheng Wang
- Shane Wesener
- Viahwakanth Potharla
- Yi-qiang Cheng
Organizations
- University of Wisconsin–Milwaukee