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CONSTRUCTION AND IDENTIFICATION OF THE CHIMERIC MS/HIL12 EUKARYOTIC EXPRESSION PLASMID.

Sen Zhong

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Abstract

Objective:To construct and identify the chimeric MS/hIL12 eukaryotic expression plasmid.Methods:Firstly MS-linker was amplified by PCR and cloned into the single Nhe I and Mil I cloning sites of pCI-neo,so pCI-neo-MS-linker(pMSL)plasmid was constructed.Secondly hIL 12 was amplified by PCR and cloned into the single Mlu I and Sal I colning sites of pMSL.Finally correct pCI-neo-MS/hIL12(pMSI)plasmid was identified by PCR,RE digestion and DNA seguencing.Results:The accuracy of pMSI plasmid construction was confirmed by a number of molecular biological technique.Conclusion:The construction of MS/hIL12 chimera by linkage of M.tuberculosis Mtb8.4 gene with the signal peptide(MS)to human IL12 gene provided the possibility for investigating a new tuberculosis vaccine and research on its immune protection.

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What this paper is about

Objective:To construct and identify the chimeric MS/hIL12 eukaryotic expression plasmid.Methods:Firstly MS-linker was amplified by PCR and cloned into the single Nhe I and Mil I cloning sites of pCI-neo,so pCI-neo-MS-linker(pMSL)plasmid was constructed.Secondly hIL 12 was amplified by PCR and cloned into the single Mlu I and Sal I colning sites of pMSL.Finally correct pCI-neo-MS/hIL12(pMSI)plasmid was identified by PCR,RE digestion and DNA seguencing.Results:The accuracy of pMSI plasmid construction was confirmed by a number of molecular biological technique.Conclusion:The construction of MS/hIL12 chimera by linkage of M.tuberculosis Mtb8.4 gene with the signal peptide(MS)to human IL12 gene provided the possibility for investigating a new tuberculosis vaccine and research on its immune protection.

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Available abstract

Objective:To construct and identify the chimeric MS/hIL12 eukaryotic expression plasmid.Methods:Firstly MS-linker was amplified by PCR and cloned into the single Nhe I and Mil I cloning sites of pCI-neo,so pCI-neo-MS-linker(pMSL)plasmid was constructed.Secondly hIL 12 was amplified by PCR and cloned into the single Mlu I and Sal I colning sites of pMSL.Finally correct pCI-neo-MS/hIL12(pMSI)plasmid was identified by PCR,RE digestion and DNA seguencing.Results:The accuracy of pMSI plasmid construction was confirmed by a number of molecular biological technique.Conclusion:The construction of MS/hIL12 chimera by linkage of M.tuberculosis Mtb8.4 gene with the signal peptide(MS)to human IL12 gene provided the possibility for investigating a new tuberculosis vaccine and research on its immune protection.

Key concepts: Plasmid, Molecular biology, Recombinant DNA, Biology, Gene, Cloning (programming), Molecular cloning, Chimera (genetics)

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CONSTRUCTION AND IDENTIFICATION OF THE CHIMERIC MS/HIL12 EUKARYOTIC EXPRESSION PLASMID. — Research Paper | ScholarLens