2008PubMedRequires access

[Construction and expression of a prokaryotic vector of recombinant human adiponectin global domain].

Pu Su, Yerong Yu, Yang Long

Open publisher page 0 citations

Abstract

OBJECTIVE: To construct and express the recombinant human adiponectin (gAd) global domain. METHODS: gAd complementary DNA (cDNA) was obtained from human fat tissue by RT-PCR. The PCR product was cloned into the vector pMD18-T and the prokaryotic expression vector pET32a(+). The recombinant vector was identified by digestion with double restriction endonucleases SalI and EcoRI, PCR and sequence analysis. The recombinant plasmid containing gAd gene was transformed into E. coli BL21 (DE3), and the expression of the fusion protein His-gAd was induced by IPTG. RESULTS: The gAd cDNA of 412 bp was obtained from the total RNA of the fat tissue and verified by sequence analysis. CONCLUSION: The recombinant plasmid could stably express the 34-kD fusion protein His-gAd in the engineered bacteria in the form of inclusion bodies.

About this research paper

What this paper is about

OBJECTIVE: To construct and express the recombinant human adiponectin (gAd) global domain. METHODS: gAd complementary DNA (cDNA) was obtained from human fat tissue by RT-PCR. The PCR product was cloned into the vector pMD18-T and the prokaryotic expression vector pET32a(+). The recombinant vector was identified by digestion with double restriction endonucleases SalI and EcoRI, PCR and sequence analysis. The recombinant plasmid containing gAd gene was transformed into E. coli BL21 (DE3), and the expression of the fusion protein His-gAd was induced by IPTG. RESULTS: The gAd cDNA of 412 bp was obtained from the total RNA of the fat tissue and verified by sequence analysis. CONCLUSION: The recombinant plasmid could stably express the 34-kD fusion protein His-gAd in the engineered bacteria in the form of inclusion bodies.

Why it matters

A significance statement is not available in the OpenAlex record.

Key contribution

A contribution statement is not available in the OpenAlex record.

Method / approach

Method details are not available in the OpenAlex metadata.

Main findings

Findings are not separately available in the OpenAlex metadata.

Limitations

Limitations are not available in the OpenAlex metadata.

Applications

Application details are not available in the OpenAlex metadata.

Available abstract

OBJECTIVE: To construct and express the recombinant human adiponectin (gAd) global domain. METHODS: gAd complementary DNA (cDNA) was obtained from human fat tissue by RT-PCR. The PCR product was cloned into the vector pMD18-T and the prokaryotic expression vector pET32a(+). The recombinant vector was identified by digestion with double restriction endonucleases SalI and EcoRI, PCR and sequence analysis. The recombinant plasmid containing gAd gene was transformed into E. coli BL21 (DE3), and the expression of the fusion protein His-gAd was induced by IPTG. RESULTS: The gAd cDNA of 412 bp was obtained from the total RNA of the fat tissue and verified by sequence analysis. CONCLUSION: The recombinant plasmid could stably express the 34-kD fusion protein His-gAd in the engineered bacteria in the form of inclusion bodies.

Key concepts: Recombinant DNA, Complementary DNA, Fusion protein, Molecular biology, Restriction enzyme, EcoRI, Biology, Plasmid

Related papers

Back to paper searchBrowse research topicsOriginal source
[Construction and expression of a prokaryotic vector of recombinant human adiponectin global domain]. — Research Paper | ScholarLens