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Constructure prokaryotic expression plasmid of KSHV K8.1 gene

Renyong Lin

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Abstract

Objective: To construct the prokaryotic expression plasmid for pET41a-K8.1. Methods: Using recombinant plasmid pGEM-Teasy/K8.1 as the template, EcoR I and Xho I restriction digestion sites were amplified by PCR and subsequently inserted into prokaryotic vector, pET-41a. The recombinant plasmid was transformed into E.coli BL21(DE3) and then identified by double enzyme digesting and sequencing. Results: The insert target DNA was 494 pb, EcoR I and Xho I restriction digestion identified that there was DNA band at about 500 pb position, sequencing showing the insert DNA was connected correctly. Conclusion: Recombinant plasmid pET41a-K8.1 is successfully constructed. Further study is needed to identify its immunity.

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What this paper is about

Objective: To construct the prokaryotic expression plasmid for pET41a-K8.1. Methods: Using recombinant plasmid pGEM-Teasy/K8.1 as the template, EcoR I and Xho I restriction digestion sites were amplified by PCR and subsequently inserted into prokaryotic vector, pET-41a. The recombinant plasmid was transformed into E.coli BL21(DE3) and then identified by double enzyme digesting and sequencing. Results: The insert target DNA was 494 pb, EcoR I and Xho I restriction digestion identified that there was DNA band at about 500 pb position, sequencing showing the insert DNA was connected correctly. Conclusion: Recombinant plasmid pET41a-K8.1 is successfully constructed. Further study is needed to identify its immunity.

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Available abstract

Objective: To construct the prokaryotic expression plasmid for pET41a-K8.1. Methods: Using recombinant plasmid pGEM-Teasy/K8.1 as the template, EcoR I and Xho I restriction digestion sites were amplified by PCR and subsequently inserted into prokaryotic vector, pET-41a. The recombinant plasmid was transformed into E.coli BL21(DE3) and then identified by double enzyme digesting and sequencing. Results: The insert target DNA was 494 pb, EcoR I and Xho I restriction digestion identified that there was DNA band at about 500 pb position, sequencing showing the insert DNA was connected correctly. Conclusion: Recombinant plasmid pET41a-K8.1 is successfully constructed. Further study is needed to identify its immunity.

Key concepts: Insert (composites), Plasmid, Recombinant DNA, Restriction enzyme, Biology, Molecular biology, DNA, Vector (molecular biology)

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