Design and Cloning of Gene Encoding SLPI C-Terminal Domain in Escherichia coli TOP10

Elevated levels of neutrophil elastase in CPOD (Chronic Obstructive Pulmonary Disease) airways are regarded as the main trigger lung destruction and inflammation. SLPI (Secretory leukocyte protease inhibitor), an inhibitor protease, represents attractive candidate for treatment chronic diseases due to proteases excess. The antiprotease active site has been located on C-terminal domain. This study aimed design clone gene encoding domain (SLPIc). SLPIc was optimized predicted solubility using OptimumGene™ SoDoPe software. nucleotide sequence synthesized, inserted into pGEX 4T-2 vector commercially by Genscript, transformed Escherichia coli TOP10. contains a glutathione S transferase (GST) before MCS generate recombinant protein fusion with GST. For purification purposes, His-tag synthesized together SLPIc. gave CAI value 0.81, GC content 52.31, CFD 2%. probability fused GST increased from 0.124 0.4656. Confirmation transformant restriction sequencing analysis showed that plasmid successfully E. TOP10 novelty this study. high being expressed based in-silico analysis.