Cloning and prokaryotic expression of human secondary lymphoid tissue chemokine

Objective:To clone and express secondary lymphoid tissue chemokine(SLC) in E.coli .Methods:SLC cDNA was amplified by RT PCR,and fragment was cloned into pMD18 T vector and then sequenced.After recombinant clone vector was digested by BamHⅠ and EcoRⅠ, the SLC purified fragment was subcloned into expression vector pBV220.The recombinant vector was induced to express SLC at 42℃.The recombinant protein was purified with molecular sieve,identified by SDS PAGE and Western blotting.Results:Both recombinant clone vector and recombinant expression vector were successfully constructed.The protein accounted for up to 15% of the whole protein of E.coli and was confirmed by Western blotting.Purity of protein was found to be above 90%.Conclusion:The recombinant human SLC is successfully expressed in prokaryotic system.