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Construction of Fusion Expression Plasmid PQE3-IL3-PE38KDEL

Author: ShenYan
Tutor: LouShiFeng
School: Chongqing Medical University
Course: Internal Medicine
Keywords: Recombinant toxin Human interleukin- 3 Pseudomonas exotoxin (PE38KDEL)
CLC: R392.1
Type: Master's thesis
Year: 2008
Downloads: 16
Quote: 0
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Abstract


Objective: To build human interleukin (IL-3) with Pseudomonas exotoxin fragment (PE38KDEL) , the fusion gene prokaryotic expression vector , and lay the foundation for further expression of the fusion protein with killing effect . Method: 1 ) extracted from human peripheral blood total RNA , IL-3 gene was amplified by RT-PCR , secondary PCR method to introduce flexible linker peptide linker fragment , PCR products were inserted into the carrier PQE30 transformed cloned enzyme digestion and DNA sequencing certainly cloning results . 2) in order to PGEX-4T-1PE38KDEL plasmid as a template , the PCR amplification PE38KDEL fragments , PCR products were inserted into the double-digested plasmid PQE30-IL-3 build PQE30 -IL - 3 - PE38KDEL , ligation products were transformed clones were digested identification and DAN sequencing. Results: In this study successfully constructed prokaryotic expression plasmids The recombinant plasmid PQE30 - IL - 3- PE38KDEL anastomosis after restriction enzyme digestion and expected results , and nucleic acid sequencing shows exactly the same sequence of insertion of the IL-3 gene and PE38KDEL gene with Genebank reported .

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