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- W2347927774 abstract "Objective To construct a prokaryotic expression vector for G13 domain of granulysin, express in E. coli BL21 (DE3) and study the effect of expressed product on the activity of host E. coli. Methods Synthesize the DNA sequence encoding G13 domain of granulysin, amplify by PCR and clone into expression vector pBAD/TOPO ThioFusion by T-A cloning. Screen positive recombinant plasmid by PCR and sequencing, and transform to E. coli BL21(DE3) for expression under induction of L-arabinose. Identify the expressed product by SDS-PAGE and determine the A600 values of host E. coli before and after induction. Test the recombinant E. coli after induction for mutant. Results The specific protein with a relative molecular mass of about 15 000 was expressed, which contained about 3% of total somatic protein. The A600 value of host E. coli decreased with the increasing time for induction, and increased gradually after expression of foreign protein. The sequencing result of extracted plasmid showed mutation at both -35 and -10 domains. The proportion of mutant cells increased with the increasing time for induction. Conclusion The prokaryotic expression vector for G13 domain of granulysin was successfully constructed, and G13 fusion protein was expressed in E. coli. The expression resulted the decrease of activity of host E. coli." @default.
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- W2347927774 date "2008-01-01" @default.
- W2347927774 modified "2023-09-28" @default.
- W2347927774 title "Expression of G13 Domain of Granulysin and Effect of Expressed Product on Activity of Host E.coli" @default.
- W2347927774 hasPublicationYear "2008" @default.
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