Domain selection combined with improved cloning strategy for high throughput expression of higher eukaryotic proteins |
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| Authors: | Yunjia Chen Shihong Qiu Chi-Hao Luan Ming Luo |
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| Affiliation: | (1) Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA;(2) Center for Biophysical Sciences and Engineering, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA |
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| Abstract: | Background Expression of higher eukaryotic genes as soluble, stable recombinant proteins is still a bottleneck step in biochemical and structural studies of novel proteins today. Correct identification of stable domains/fragments within the open reading frame (ORF), combined with proper cloning strategies, can greatly enhance the success rate when higher eukaryotic proteins are expressed as these domains/fragments. Furthermore, a HTP cloning pipeline incorporated with bioinformatics domain/fragment selection methods will be beneficial to studies of structure and function genomics/proteomics. |
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