人白细胞介素-24基因的克隆及序列测定

Cloning and Sequencing of cDNA Encoding Human Interleukin-24

目的 克隆人白细胞介素 - 2 4 (hIL - 2 4 )基因 ,以供重组表达。方法 利用自行设计合成的一对引物 ,采用RT -PCR技术从LPS活化的人外周血单个核细胞的总RNA中分离hIL - 2 4cDNA并重组到 pUC19载体上 ,进行酶切鉴定和DNA序列测定。结果 PCR及酶切产物电泳结果均证明所克隆的基因为 6 2 1bp。经序列测定证实所克隆的hIL - 2 4与GenBank报道的结果完全一致。结论 该基因系国内首次获得并经序列测定证实的hIL - 2 4cDNA基因。这为进一步在真核或大肠杆菌中高效表达有活性的重组hIL - 2 4 ,更深入地研究其作用机制以及临床应用奠定了坚实的基础。

Objective Extract gene fragment of hIL-24 for further recombinant expression. Methods cDNA fragment encoding hIL-24 was amplified from LPS-stimulated human peripheral blood mononuclear cells by reverse transcription(RT)-PCR and inserted into pUC19 plasmid,identified by endonuclease digestion and DNA sequencing. Results cDNA encoding human interleukin-24 was cloned into cloning vector pUC19 and verified by endonuclease digestion and sequencing. Conclusion This is the first time for us to get the human IL-24 cDNA clone in China. This study paves the way for future research in expression of hIL-24 in eukaryotic cells and E.coli . and in mechanism of hIL-24 action and clinical application of hIL-24.