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[Structure and regulative function of the 5' end flanking sequence of gene CCR5]
Xin-ping Sha1, Guo-ling Hu, Yu-tao Xie
1Institute of Infectious Disease, Xiangya Hospital, Central South University, Changsha 410008, China.
Objective:
To construct the pCAT reporter genes containing the 5'-end flanking of CC chemokine receptor 5(CCR5) gene in different sequence lengths and identify the sequence, which regulates the gene expression of CCR5 by the CAT analysis system.
Methods:
The target sequences were amplified by pyrobest DNA polymerase, and were inserted into the upstream of CAT gene located in the pCAT enhancer vector by the directional clone technique respectively; the regulative sequence was identified by analyzing the CAT activities of reconstructed plasmid in Hela cells.
Results:
The region, containing 486 bp upstreaming from exon 1, stimulated the reporter gene activity which was about 3 times that of the pCAT enhancer vector in transfected cells.
Conclusion:
There is an up-regulative element of gene transcription in the region of -1(-)-486 bp in CCR5 gene upstream.