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- W2368907998 abstract "Objective: To design and synthesize codon-optimized MoPn-maior outer membrane protein (MOMP) gene(HuMOMP),construct recombinant eukaryotic expression plasmids of HuMOMP gene and EGFP-HuMOMP fusion gene. Methods: The differences of preferable codon usage between chlamydia muridarum(MoPn) and mammal gene were analyzed,and the sequence of codon-optimized MoPn-MOMP (HuMOMP) was designed and synthesized. Plasmid pUC-HuMOMP was digested with KpnI and EcoRI to get HuMOMP segment,which was cloned into vector pcDNA3 to construct pcDNA3-HuMOMP. Plasmid pcDNA3-HuMOMP was digested with KpnI and ApaI to get HuMOMP segment,which was cloned into vector pEGFP-C1 to construct pEGFP-HuMOMP. After confirming its correctness by restriction enzyme analysis,pEGFP-HuMOMP was transfected into COS-1 cell instantly with liposome,and observed its expression using fluorescence microscope after 24 hours. Results: Codon-optimized MoPn-MOMP gene (HuMOMP) was designed and synthesized,then verified by gene sequencing. The recombinant plasmids pcDNA3- HuMOMP and pEGFP-HuMOMP digested with corresponding restriction enzymes could get correct DNA segments. EGFP -HuMOMP fusion gene was expressed effectively in COS-1 cells. Fluorescent protein mainly existed in cytoplasm,which further certified the sequence precision of HuMOMP gene and the constructive correction of fluorescent expression plasmids. Conclusion: Designed and synthesized codon-optimized MoPn-MOMP gene (HuMOMP) can be expressed in mammalian cell successfully." @default.
- W2368907998 created "2016-06-24" @default.
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- W2368907998 date "2005-01-01" @default.
- W2368907998 modified "2023-09-23" @default.
- W2368907998 title "Design and expression of codon optimized chlamydia trachomatis major outer membrane protein gene" @default.
- W2368907998 hasPublicationYear "2005" @default.
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