2004Anhui Nongye Daxue xuebaoRequires access

Cloning of Chicken Interleukin-2 Gene and Constructionand Expression of the Recombinant Plasmid

Xu FaZhi, Jinnian Li

Open publisher page 0 citations

Abstract

In this study,a cDNA fragment of chicken IL-2 about 439 bp was cloned from ConA- stimulated chicken spleen cells by reverse transcription (RT)-PCR with a pair of designed primers. Then it was inserted into pcDNA3, an eukaryotic expression plasmid by a directional link, identified by endonuclease and DNA sequencing, which conformed to chicken IL-2 gene fragment reported in the literature. This fragment was further inserted into a prokaryotic expressive plasmid pGEX-4T-1, and then it was transferred into the strain BL21. After the bacteria were induced and cultured, expressed protein was separated. A band with 42 000 of molecular weight was observed on SDS-PAGE. All these results suggest that the cloned gene of chicken IL-2 is expressed in prokaryotic cells.

About this research paper

What this paper is about

In this study,a cDNA fragment of chicken IL-2 about 439 bp was cloned from ConA- stimulated chicken spleen cells by reverse transcription (RT)-PCR with a pair of designed primers. Then it was inserted into pcDNA3, an eukaryotic expression plasmid by a directional link, identified by endonuclease and DNA sequencing, which conformed to chicken IL-2 gene fragment reported in the literature. This fragment was further inserted into a prokaryotic expressive plasmid pGEX-4T-1, and then it was transferred into the strain BL21. After the bacteria were induced and cultured, expressed protein was separated. A band with 42 000 of molecular weight was observed on SDS-PAGE. All these results suggest that the cloned gene of chicken IL-2 is expressed in prokaryotic cells.

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

In this study,a cDNA fragment of chicken IL-2 about 439 bp was cloned from ConA- stimulated chicken spleen cells by reverse transcription (RT)-PCR with a pair of designed primers. Then it was inserted into pcDNA3, an eukaryotic expression plasmid by a directional link, identified by endonuclease and DNA sequencing, which conformed to chicken IL-2 gene fragment reported in the literature. This fragment was further inserted into a prokaryotic expressive plasmid pGEX-4T-1, and then it was transferred into the strain BL21. After the bacteria were induced and cultured, expressed protein was separated. A band with 42 000 of molecular weight was observed on SDS-PAGE. All these results suggest that the cloned gene of chicken IL-2 is expressed in prokaryotic cells.

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

Related papers

Back to paper searchBrowse research topicsOriginal source
Cloning of Chicken Interleukin-2 Gene and Constructionand Expression of the Recombinant Plasmid — Research Paper | ScholarLens