[Cloning, sequencing and in vitro expression of human RANTES gene].
An expected 276 bp fragment of the gene precursor encoding the signal peptide and mature protein of human beta-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.
Zeng
Q P
QP
Tropical Medicine Institute, Guangzhou University of Traditional Chinese Medicine, Guangzhou 510405, China. qpzeng@usa.net
Yang
R Y
RY
Feng
L L
LL
Fu
L C
LC
D000163:Acquired Immunodeficiency Syndrome / Q000628:therapy; D001483:Base Sequence; D018946:Chemokine CCL5 / Q000737:chemistry / Q000235:genetics*; D003001:Cloning, Molecular; D015316:Genetic Therapy; D006801:Humans; D008969:Molecular Sequence Data
D018946:Chemokine CCL5