An expected 276bp fragment of the gene precursor encoding the signal peptide and mature protein of human β chemokine RANTES was amplified by reverse transcription\|polymerase chain reaction (RT\|PCR) from RNA of PHA\...An expected 276bp fragment of the gene precursor encoding the signal peptide and mature protein of human β chemokine RANTES was amplified by reverse transcription\|polymerase chain reaction (RT\|PCR) from RNA of PHA\|activated human peripheral blood lymphocytes.This putative interested gene was inserted directly into a T\|vector and the ligation was confirmed by restriction enzyme digestion.The sequence data of the cloned fragment showed that it was almost identical with published sequences of RANTES gene,except for only one nucleotide substitution within the signal peptide region.The \%in vitro\% expressed recombinant RANTES protein was detected by the chemiluminescence enzyme\|linked immune Dot blotting assay after combining the recombinant plasmid with the \%in vitro\% SP6/T7 transcription and translation system.The successful cloning and expression of RANTES gene should shed light on future′s gene therapy of AIDS.展开更多
文摘An expected 276bp fragment of the gene precursor encoding the signal peptide and mature protein of human β chemokine RANTES was amplified by reverse transcription\|polymerase chain reaction (RT\|PCR) from RNA of PHA\|activated human peripheral blood lymphocytes.This putative interested gene was inserted directly into a T\|vector and the ligation was confirmed by restriction enzyme digestion.The sequence data of the cloned fragment showed that it was almost identical with published sequences of RANTES gene,except for only one nucleotide substitution within the signal peptide region.The \%in vitro\% expressed recombinant RANTES protein was detected by the chemiluminescence enzyme\|linked immune Dot blotting assay after combining the recombinant plasmid with the \%in vitro\% SP6/T7 transcription and translation system.The successful cloning and expression of RANTES gene should shed light on future′s gene therapy of AIDS.
