Herein is proposed an strategy for expanding the genetic alphabet via engineering unnatural base pair acceptance by DNA polymerases. This research proposal addresses the following specific aims: 1) to engineer a DNA polymerase that processes unnatural base pairs with high fidelity, 2) to identify the structure-function relationships that dictate polymerase fidelity and processivity, and 3) to clarify the role of hydrogen bonding with respect to base pair stability. The incorporation of any of the proposed base pairs will create 152 additional codons that can be applied to broader studies of protein structure and function within the Schultz group. This effort will provide unprecedented access to the incorporation of multiple unnatural amino acids into a single protein sequence, the implications of which are numerous. The successful incorporation of unnatural base pairs by engineered DNA polymerases will represent a landmark achievement and will significantly advance the frontiers of science and medicine.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
5F32GM019833-02
Application #
6138319
Study Section
Bio-Organic and Natural Products Chemistry Study Section (BNP)
Program Officer
Marino, Pamela
Project Start
1999-12-16
Project End
Budget Start
1999-12-16
Budget End
2000-04-28
Support Year
2
Fiscal Year
2000
Total Cost
$14,081
Indirect Cost
Name
Scripps Research Institute
Department
Type
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Berger, M; Wu, Y; Ogawa, A K et al. (2000) Universal bases for hybridization, replication and chain termination. Nucleic Acids Res 28:2911-4