Keywords

Restriction Display, Linker Design, Peptide Library

Abstract

In order to explore the flexible linker design of restriction display (rd) technology in constructing protein peptide libraries, three sets of linkers were designed based on the prokaryotic expression vector pet22b and the yeast expression vector pnmt-topo. The three sets of linkers were increased by one base in order to ensure It is always possible for the connected fragments to express the correct open reading frame. Then, using the complete gene plasmid DNA of the HIV-1b subtype representative strain U26942 as the object, the corresponding protein peptide libraries were established using rd technology. Randomly select from each library Twelve clones were selected for sequencing analysis and protein expression prediction. As a result, a clone that could express hivpol polypeptide was obtained from the prokaryotic expression library. The sds-polyacrylamide gel electrophoresis (sds-page) results showed that the clone was in bacteria bl21 ( de3) has higher expression, and the Western blot is positive, consistent with theoretical predictions. These results suggest that rd technology is a new method for establishing genomic random peptide libraries. The flexible linker design of this method can meet the needs of different expression vectors. .

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Original research done by Ma Wenli, Liu Jia, Li Ling, Zhang Bao, Shi Yongxia, Zheng Wenling

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