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A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells
Published on: July 31, 2012
[Construction of recombinant plasmids containing genes of HBsAg and their expression in the eukaryotic cells]
1Viral Hepatitis Research Unit, First Affiliated Hospital, WCUMS, Chengdu 610041, China.
Objective:
To construct recombinant plasmids expressing L, M, S and pre-S1-S protein of HBsAg.
Methods:
Amplifying segments of S, pre-S2-S, pre-S1-pre-S2-S genes of HBV by PCR and amplifying segment of pre-S1-S by overlap extension PCR; inserting the segments into Rc/CMV and pSG5UTPL/Flag plasmids respectively and exploring their expressions by Western-Blot hybridization, identifying the inserting segments by sequencing.
Results:
The sequences of the inserted segments were the same as the genes of S, pre-S2-S, pre-S1-pre-S2-S and pre-S1-S and the results of Western-Blot hybridization were positive for the aimed proteins.
Conclusion:
We have gained 8 recombinant plasmids expressing S, M, L and pre-S1-S proteins with high efficacy.
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