Molecular cloning of rat interleukin-10 cDNA

Objective To clone the full length of rat interleukin-10 cDNA as a piece of pioneer work for its use in molecular biology.?Methods The rat spleen was removed to harvest the spleenocytes under sterile condition. Having been cultured under the stimulation of LPS for 4 hours, the spleenocytes were collected and used for extraction of total RNA. The cDNA of rat IL-10 was cloned from RNA by RT-PCR and subcloned into pcDNA 3 vectors. By transfering to compentent cells, the recombinant pcDNA 3 IL (encoding IL-10) vectors were prepared for identification and sequencing.?Results The transcription of IL-10 was increased in spleenocytes after being stimulated by LPS, and the expected band was readily produced by RT-PCR. It was confirmed that the sequence of rat IL-10 cDNA cloned was identical to what is reported by Genbank.?Conclusion The full-length sequence of rat IL-10 cDNA is successfully cloned.