2006Journal of Jilin UniversityRequires access

Construction and identification of human decorin gene recombinant eukaryotic expressing vector

Hai-Ping Cao, SU Zhen-bo

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Abstract

Objective To construct a recombinant eukaryotic expressing vector pcDNA-dec and provide a basis for further study on the bioactivity of decorin(DCN).Methods DCN cDNA was amplified by using polymerase chain reaction(PCR).Product of PCR and expressing vector were digested by restriction endonucleases,then ligated and transformed into JM109 bacteria.Results The specific DNA fragment was obtained by PCR as supposed.Product of PCR and expressing vector were digested by restriction endonucleases,then ligated to establish the recombinant eukaryotic expression vector pcDNA-dec.It was confirmed that DCN cDNA was inserted into the eukaryotic expression vector correctly by using digestion identification and sequencing.Conclusion The recombinant eukaryotic expression vector pcDNAdec of human DCN is successfully constructed.

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Objective To construct a recombinant eukaryotic expressing vector pcDNA-dec and provide a basis for further study on the bioactivity of decorin(DCN).Methods DCN cDNA was amplified by using polymerase chain reaction(PCR).Product of PCR and expressing vector were digested by restriction endonucleases,then ligated and transformed into JM109 bacteria.Results The specific DNA fragment was obtained by PCR as supposed.Product of PCR and expressing vector were digested by restriction endonucleases,then ligated to establish the recombinant eukaryotic expression vector pcDNA-dec.It was confirmed that DCN cDNA was inserted into the eukaryotic expression vector correctly by using digestion identification and sequencing.Conclusion The recombinant eukaryotic expression vector pcDNAdec of human DCN is successfully constructed.

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Available abstract

Objective To construct a recombinant eukaryotic expressing vector pcDNA-dec and provide a basis for further study on the bioactivity of decorin(DCN).Methods DCN cDNA was amplified by using polymerase chain reaction(PCR).Product of PCR and expressing vector were digested by restriction endonucleases,then ligated and transformed into JM109 bacteria.Results The specific DNA fragment was obtained by PCR as supposed.Product of PCR and expressing vector were digested by restriction endonucleases,then ligated to establish the recombinant eukaryotic expression vector pcDNA-dec.It was confirmed that DCN cDNA was inserted into the eukaryotic expression vector correctly by using digestion identification and sequencing.Conclusion The recombinant eukaryotic expression vector pcDNAdec of human DCN is successfully constructed.

Key concepts: Recombinant DNA, Complementary DNA, Restriction enzyme, Vector (molecular biology), Molecular biology, Biology, Polymerase chain reaction, Expression vector

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